05_eph_4.1-7_wayne_seaton

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Chapter 7. Nucleic Acid1. Definition(1). Cyclic nucleotides(2). Chargaff’s Rule(3). Double Helix(4). B- form DNA and Z-form DNA(5). 5’-Cap of mRNA(6). Denaturation and renaturation(7). Tm2. Mono-choice questions(1) The compound that consists of ribose linked by an N-glycosidic bond to N-9 of adenine is:A. a deoxyribonucleoside.B. a purine nucleotide.C. a pyrimidine nucleotide.D.adenosine monophosphate.E.adenosine.(2) A major component of RNA but NOT of DNA is:A. adenine.B.cytosine.C.guanine.D.thymine.E.uracil.(3) The difference between a ribonucleotide and a deoxyribonucleotide is:A. a deoxyribonucleotide has an —H instead of an —OH at C-2.B. a deoxyribonucleotide has α configuration; ribonucleotide has the β configuration atC-1.C. a ribonucleotide has an extra —OH at C-4.D. a ribonucleotide has more structural flexibility than deoxyribonucleotide.E. a ribonucleotide is a pyranose, deoxyribonucleotide is a furanose.(4) The phosphodiester bonds that link adjacent nucleotides in both RNA and DNA:A.always link A with T and G with C.B.are susceptible to alkaline hydrolysis.C.are uncharged at neutral pH.D.form between the planar rings of adjacent bases.E.join the 3' hydroxyl of one nucleotide to the 5' hydroxyl of the next.(5) The DNA oligonucleotide abbreviated pATCGAC:A.has 7 phosphate groups.B.has a hydroxyl at its 3' end.C.has a phosphate on its 3' end.D.has an A at its 3' end.E.violates Chargaff's rules.(6) The experiment of Avery in which nonvirulent bacteria were made virulent by transformation was significant because it showed that:A.bacteria can undergo transformation.B.genes are composed of DNA only.C.mice are more susceptible to pneumonia than are humans.D.pneumonia can be cured by transformation.E.virulence is determined genetically.(7) Chargaff's rules state that in typical DNA:A. A = G.B. A =C.C. A = U.D. A + T = G + C.E. A + G = T + C.(8) Based on Chargaff's rules, which of the following are possible base compositions for double-stranded DNA?%A %G %C%T %UA. 5 45 45 5 0B. 20 20 20 20 20C. 35 15 35 15 0D.All of the above.E.None of the above.(9) In the Watson-Crick model of DNA structure:A.both strands run in the same direction, 3' 5'; they are parallel.B.phosphate groups project toward the middle of the helix, where they are protectedfrom interaction with water.C.T can form three hydrogen bonds with either G or C in the opposite strand.D.the distance between the sugar backbone of the two strands is just large enough toaccommodate either two purines or two pyrimidines.E.the distance between two adjacent bases in one strand is about 3.4 Å.(10) Which of the following is NOT true of all naturally occurring DNA?A.Deoxyribose units are connected by 3',5'-phosphodiester bonds.B.The amount of A always equals the amount of T.C.The ratio A+T/G+C is constant for all natural DNAs.D.The two complementary strands are antiparallel.E.Two hydrogen bonds form between A and T.(11) In the Watson-Crick model of DNA structure (now called B-form DNA):A. a purine in one strand always hydrogen bonds with a purine in the other strand.B.A–T pairs share three hydrogen bonds.C.G–C pairs share two hydrogen bonds.D.the 5' ends of both strands are at one end of the helix.E.the bases occupy the interior of the helix.(12) The double helix of DNA in the B-form is stabilized by:A.covalent bonds between the 3' end of one strand and the 5' end of the other.B.hydrogen bonding between the phosphate groups of two side-by-side strands.C.hydrogen bonds between the riboses of each strand.D.nonspecific base-stacking interaction between two adjacent bases in the same strand.E.ribose interactions with the planar base pairs.(13) B-form DNA in vivo is a ________-handed helix, _____ Å in diameter, with a rise of ____ Å per base pair.A.left; 20; 3.9B.right; 18; 3.4C.right; 18; 3.6D.right; 20; 3.4E.right; 23; 2.6(14) In double-stranded DNA:A.only a right-handed helix is possible.B.sequences rich in A–T base pairs are denatured less readily than those rich in G–Cpairs.C.the sequence of bases has no effect on the overall structure.D.the two strands are parallel.E.the two strands have complementary sequences.(15) Which of the following is a palindromic sequence?A.AGGTCCTCCAGGTTCCGCAAGGC.GAATCCCTTAGGD.GGATCCCCTAGGE.GTA TCCCATAGG(16) Which of the following are possible base compositions for single-stranded RNA?%A %G %C%T%UA. 5 45 45 0 5B. 25 25 25 0 25C. 35 10 30 0 25D.All of the above.E.None of the above.(17) Double-stranded regions of RNA:A.are less stable than double-stranded regions of DNA.B.can be observed in the laboratory, but probably have no biological relevance.C.can form between two self-complementary regions of the same single strand of RNA.D.do not occur.E.have the two strands arranged in parallel (unlike those of DNA, which areantiparallel).(18) When double-stranded DNA is heated at neutral pH, which change does not occur?A.The absorption of ultraviolet (260 nm) light increases.B.The covalent N-glycosidic bond between the base and the pentose breaks.C.The helical structure unwinds.D.The hydrogen bonds between A and T break.E.The viscosity of the solution decreases.(19) Which of the following deoxyoligonucleotides will hybridize with a DNA containing the sequence 5'AGACTGGTC3' ?A.5'CTCA TTGAG3'B.5'GACCAGTCT3'C.5'GAGTCAACT3'D.5'TCTGACCAG3'E.5'TCTGGA TCT3'(20) The ribonucleotide polymer 5'GTGATCAAGC3' could only form a double-stranded structure with:A.5'CACTAGTTCG3'.B.5'CACUAGUUCG3'.C.5'CACUTTCGCCC3'.D.5'GCTTGA TCAC3'.E.5'GCCTAGTTUG3'.(21) In the laboratory, several factors are known to cause alteration of the chemical structure of DNA. The factor(s) likely to be important in a living cell is (are):A.heat.B.low pH.C.oxygen.D.UV light.E.both C and D.(22) In living cells, nucleotides and their derivatives can serve as:A.carriers of metabolic energy.B.enzyme cofactors.C.intracellular signals.D.precursors for nucleic acid synthesis.E.all of the above.(23) ATP is NOT a nucleoside because it ________.A. has phosphate groupsB. has three phosphates instead of just oneC. lacks the deoxyribosyl groupD. is not connect to a carbohydrate group(24) According to Chargaff’s observations of nucleotide composition of DNA samplesA.% of (G + C) + % of (A + T) = 100%.B.A = T.C.G = C.D.%A + %G + %C + %T = 100%.E.All of the above(25) The rise and pitch of B-DNA are 0.33 nm and 3.40 nm, respectively. About how many helical urns are there in a fragment 1 mm in length?A. 3030B. 294C. 330D. 0.0034E. Cannot calculate from the information given.(26) Regions of DNA that are most easily unwound haveA. about half G and half C.B. alternating A and G.C. greater G:C content.D. greater A:T content.(27) Which is NOT true of the different conformations of DNA?A. Z-DNA is a left-handed spiral.B. A-DNA and B-DNA are right-handed spirals.C. A-DNA and Z-DNA segments are limited to short regions of DNA.D. Both A-DNA and B-DNA are dehydrated.(28) In addition to knowing the chemical structures of the nucleotides, Watson and Crick used________ of Franklin and Wilkins and the chemical equivalencies of Chargaff in order to propose their model of DNA structure.A. sequence informationB. UV spectraC. % (G + C) and % (A + T)D. X-ray diffraction data(29) In proteins, amino acids are linked by peptide bonds; in polynucleotides, nucleotides are linked byA. phosphoanhydride bonds.B. 3’-5’phosphodiester bonds.C. 5’-3’phosphodiester bonds.D. B and CE. All of the above(30) It is easier to melt DNA richer in AT than GC becauseA. it is more heat sensitive.B. there is one less hydrogen bond.C. the helix pitch is longer in AT rich regions.D. All of the above(31) As B-DNA is gradually heated, the absorbance at 260 nmA. increases.B. decreases.C. stays the same.D. is half way between that of poly (AT and poly (GC).(32) Which of the following is mismatched?A. rRNA: 80% of cellular RNAB. tRNA: carry amino acids during protein synthesisC. mRNA: stable RNA carrying the coded information from DNAD. small RNA: catalytic with or without proteins(33) Which type of RNA is the most abundant in living cells (by percent)?A. ribosomalB. messengerC. smallD. transfer(34) Which is NOT a difference between RNA and DNA?A. The sugar ring of RNA is more oxidized than that in DNA.B. RNA contains uracil; DNA usually does not.C. RNA cannot form helices.D. RNA is single-stranded; DNA is double-stranded.(35) How does a nucleotide differ from a nucleoside?A. Nucleosides are found in DNA, whereas nucleotides are found in RNA.B. Purines are only found in nucleotides.C. Nucleosides contain only deoxyribose sugars.D. A nucleotide is a nucleoside with a phosphate ester linked to the sugar .E. None of the above.(36) The feature(s) of DNA deduced by Watson and Crick includedA. two antiparallel polynucleotide chains coiled in a helix around a common axis.B. the pyrimidine and purine bases lie on the inside of the helix.C. the bases are nearly perpendicular to the axis.D. All of the above.E. None of the above.(37) The chemical forces that contribute to the stability of the DNA due to the base stacking present in the DNA helix areA. hydrogen bonds.B. van der Waals.C. disulfide bonds.D. B and C.E. None of the above.3. Short answer questions(1). A viral DNA is analyzed and found to have the following base composition, in mole percent: A = 32, G = 16, T = 40, C = 12.A. What can you immediately conclude about this DNA?B. What kind of secondary structure do you think it would have?(2). Give the following sequence for one strand of a double-strand oligonucleotide:5’ ACCGTAAGGCTTTAG 3’A. Write the sequence for the complementary DNA strand.B. Write the sequence of the RNA complementary to the strand shown above.(3). A stretch of double-stranded DNA contains 1000 bp, and its base composition is 58%(G+C). How many thymine residues are in this region of DNA?(4). Do the two complementary strands of a segment of DNA have the same base composition?Does (A+G) equal (C+T)?(5). In samples of DNA isolated from two unidentified species of bacteria, X and Y, adenine makes up 32% and 17%, respectively, of the total bases. What relative proportions of adenine, guanine, thymine, and cytosine would you expect to find in the two DNA samples? What assumptions have you made? One of these species was isolated from a hot spring (64℃). Suggest which species is the thermophilic bacterium. What is the basis for your answer?(6). Calculate the weight in grams of a double-helical DNA molecule stretching from the earth to the moon (~320,000 km). The DNA double helix weighs about 1 X 1018 g per 1,000 nucleotide pairs; each base pair extends 3.4 Å. For an interesting comparison, your body contains about 0.5 g of DNA!(7). Compare hydrogen bonding in the αhelix of proteins and in the double helix of DNA. Include the answer the role of hydrogen bonding in stabilizing these two structures.(8). Describe qualitatively how the t m for a double-stranded DNA depends upon its nucleotide composition.(9). Write the structure of cAMP and cGMP molecules.。

VetMAX M. bovis 核苷酸纯化协议说明书

VetMAX™ M. bovis KitNucleic acid purification protocols optimized for use with the kit (Cat. No. MPBO50) Pub. No. MAN0019166 Rev. A.0WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. Safety Data Sheets (SDSs) are available from /support.WARNING! BIOHAZARD. Read the biological hazard safety information at this product’s page at . Follow all applicable local, state/provincial, and/or national regulations for working with biological samples.■Purpose of this guide (1)■Sample selection (2)■Sample storage (2)■Required materials not supplied (2)■Purify DNA using the MagMAX™ CORE Nucleic Acid Purification Kit (automated method) (5)■Prepare samples for purification with other kits (9)■Purify DNA using the MagVet™ Universal Isolation Kit (automated method) (10)■Purify DNA using the QIAamp™ DNA Mini Kit (manual method) (11)■Purify DNA using the NucleoSpin™ Tissue kit (manual method) (13)■Good laboratory practices for PCR and RT-PCR (14)Appendix A Purification with the KingFisher™ Duo Prime or KingFisher™ mL instrument■Required materials not supplied (14)■Purification procedure (15)Appendix B Documentation and support■Customer and technical support (15)■Limited product warranty (16)Purpose of this guideThis guide describes Mycoplasma bovis DNA purification protocols that have been validated and optimized for downstream use with the Applied Biosystems™ VetMAX™ M. bovis Kit (Cat. No. MPBO50).•Automated nucleic acid purification is performed using one of the following instruments: KingFisher™ Flex, MagMAX™ Express-96, KingFisher™ mL, or KingFisher™ Duo Prime.•Manual nucleic acid purification uses silica-based spin columns.Sample selectionSample storageRequired materials not suppliedUnless otherwise indicated, all materials are available through . "MLS" indicates that the material is available from or another major laboratory supplier.Materials required for sample collection, preparation, and nucleic acid purificationTable 1 Materials required for all sample preparation methodsTable 2 Additional materials required for purification from organ samplesAdditional materials required for automated nucleic acid purification Table 3 Materials required for the MagMAX™ CORE Nucleic Acid Purification KitTable 4 Materials required for the MagVet™ Universal Isolation KitAdditional materials required for manual nucleic acid purificationPurify DNA using the MagMAX™ CORE Nucleic Acid Purification Kit (automated method)Follow this procedure if you are using these instruments:•KingFisher™ Flex•MagMAX™ Express-96Follow Appendix A, “Purification with the KingFisher™ Duo Prime or KingFisher™ mL instrument” if you are using these instruments:•KingFisher™ Duo Prime•KingFisher™ mLWorkflowProcedural guidelines•Before use, invert bottles of solutions and buffers to ensure thorough mixing.•To prevent cross-contamination:–Cover the plate or tube strip during the incubation and shaking steps, to prevent spill-over.–Carefully pipet reagents and samples, to avoid splashing.•To prevent nuclease contamination:–Wear laboratory gloves during the procedures. Gloves protect you from the reagents, and they protect the nucleic acid from nucleases that are present on skin.–Use nucleic acid-free pipette tips to handle the reagents, and avoid putting used tips into the reagent containers.–Decontaminate lab benches and pipettes before you begin.Before first use of the kit(Optional) Determine the optimal bead mill homogenizer settingsWe recommend using the Fisher Scientific™ Bead Mill 24 Homogenizer for maximum nucleic acid yield. If an alternative instrument is used, follow the manufacturer's guidelines to determine the speed and time settings necessary to achieve sufficient cell lysis.Download and install the scriptThe appropriate script for the MagMAX ™CORE Nucleic Acid Purification Kit must be installed on the instrument before first use.1.On the MagMAX ™CORE Nucleic Acid Purification Kit product web page (at , search by catalogue number), scrollto the Product Literature section.2.Right ‑click the appropriate file to download the latest version of the MagMAX_CORE script for your instrument.Table 5 Recommended scriptsIf required by your laboratory, use one of the following scripts, which do not heat the samples during the elution step.Table 6 Alternate scripts without heated elution step3.See your instrument user guide or contact Technical Support for instructions for installing the script.Perform the purification procedurea.Set up the processing plates.Table 7 Plate setup: KingFisher ™ Flex or MagMAX ™ Express-96 instrument[1]Position on the instrument.b.(Optional ) To prevent evaporation and contamination, cover the prepared processing plates withsealing foil until they are loaded into the instrument.1Set up the processing platesPrepare samples as described.[1]Select the preparation method that is appropriate for your laboratory.2Prepare the sample Calculate the number of samples. Scale the components proportionally based on the volume per sample, then add 10% overage.a.For each sample, combine the following components as indicated.IMPORTANT! Add the components in the order indicated at the time of use; do not mix inadvance.bine the MagMAX ™CORE Lysis Solution with PBS (1X), pH 7.4.2.Invert the tube several times to mix, then centrifuge briefly to collect contents at the bottomof the tube.3Prepare the Lysis/PK Solution3Prepare the Lysis/PK Solution (continued)3.Add MagMAX ™CORE Proteinase K to the diluted Lysis Solution.Note: PK Buffer is not required for this protocol.b.Invert the tube several times to mix, then centrifuge briefly to collect contents at the bottom ofthe tube.Perform this procedure in single tubes – do not use plates – to avoid bine the following components in the order indicated.b.Vortex briefly to mix the sample with the Lysis/PK Solution.c.d.Centrifuge briefly to collect contents at the bottom of the tube.4Treat samples with the Lysis/PK Solutiona.Vortex the MagMAX ™CORE Magnetic Beads thoroughly to ensure that the beads are fullybine the following reagents in the order indicated.Table 8 Final Sample Plate volumes: KingFisher ™ Flex or MagMAX ™ Express-96 instrument[1]Position on the instrument.c.Immediately proceed to process samples on the instrument (next section).5Combine samples with the binding solution andbeadsa.Select the appropriate script on the instrument (see “Download and install the script” on page 6).b.Start the run, then load the prepared plates in the appropriate positions when prompted by theinstrument.6Process samples on the instrument6Process samples on the instrument(continued)Store purified nucleic acid on ice for immediate use, at −20°C for up to 1 month, or at −80°C for long‑term storage.Prepare samples for purification with other kits Prepare samples as described.Purify DNA using the MagVet ™ Universal Isolation Kit (automated method)The following protocol can be used with the KingFisher ™Flex, KingFisher ™mL, and MagMAX ™Express-96 instruments.Before first use of the kitNote: PK and MBL2 Buffer must be ordered separately from the kit.•Prepare the NM1 Buffer—Transfer 100 mL of N1 Buffer to the bottle of M1 Buffer (25 mL), then vortex to mix thoroughly.Store the NM1 Buffer at room temperature for up to 1 year.•Reconstitute the PK—Follow the recommendations of the supplier.Before each use of the kitPrepare MBL2+Beads Mix—Combine the following components for the required number of samples plus 5–10% overage, then vortex to mix thoroughly.Discard the MBL2+Beads Mix after use.Perform the purification procedurebine the following components in the order indicated, then homogenize the sample.b.Incubate at 70°C for 30 minutes.1Treat the lysate with PK Set up the processing plates or strips outside the instrument as described in the following table.[1]Position on the instrument.[2]Does not apply if using tube strips.2Set up the processing plates or stripsa.When the 70°C incubation is complete, centrifuge the samples briefly to bring downcondensation.b.Transfer the entire sample lysate to the appropriate wells in position 1 of the strip or plate 1,depending on the instrument used.c.Vortex the MBL2+Beads Mix thoroughly to ensure that the beads are fully resuspended.d.Add 520 µL of MBL2+Beads Mix to each sample and control.e.Select the appropriate script on the instrument.•KingFisher ™mL: NM_LSI_15prep•KingFisher ™Flex/MagMAX ™Express-96: NM_LSI_RRC96f.Start the run, then load the prepared processing plates or strips in their positions when prompted by the instrument.g.Load the plate or strip containing the samples and controls at position 1 when prompted by theinstrument.h.At the end of the run, when prompted by the instrument, remove the plate or tubes containingStore the purified nucleic acid at 2–8°C for immediate use or below –16°C for long-term storage.3Process samples on the instrumentPurify DNA using the QIAamp ™ DNA Mini Kit (manual method)Before first use of the kit•Reconstitute the AW1 and AW2 Buffer—Add the required volume of 96–100% ethanol according to the recommendations of the supplier.Perform the purification procedurebine the following components in the order indicated, then immediately proceed to the nextstep.b.Vortex for 15 seconds.c.Incubate at 70℃ for 30 minutes.d.Allow the tubes to cool, then centrifuge the samples briefly to bring down condensation.e.Add 200 μL of AL Buffer, then vortex for 15 seconds.1Lyse, then homogenize the samples1Lyse, thenhomogenize thesamples (continued)f.Incubate at 70℃ for 10 minutes.g.Allow the tubes to cool, then centrifuge briefly.h.Add 200 μL of 96–100% ethanol to each sample, vortex for 15 seconds, then briefly centrifugeto collect the contents.a.Insert a QIAamp™ DNA Mini Kit column into a collection tube, then transfer the entire samplevolume to the column.b.Cap the column, then centrifuge the assembly at 15,000 × g for 1 minute.c.Discard the collection tube, then place the column on a new collection tube.2Bind the DNA to thecolumna.Add 500 μL of AW1 Buffer to each column, cap the column, then centrifuge at 15,000 × g for1 minute.b.Discard the collection tube, then place the column on a new collection tube.c.Add 500 μL of AW2 Buffer to each column, cap the column, then centrifuge at 15,000 × g for1 minuted.Discard the collection tube, then place the column on a new collection tube.e.Centrifuge at 15,000 × g for 3 minutes to dry the membrane.f.Discard the collection tube.g.Place the column on a new 1.5‑mL microtube, then add 200 μL of AE Buffer.h.Cap the column, then incubate at room temperature for 1 minute.i.Centrifuge at 6,000 × g for 1 minute, then discard the column.The purified DNA is in the microtube.Store the purified DNA at 2–8°C for immediate use or below –16°C for long-term storage.3Wash, then elute the DNAPurify DNA using the NucleoSpin ™ Tissue kit (manual method)Before first use of the kit•Reconstitute the B5 Buffer—Add the required volume of 96–100% ethanol according to the recommendations of the supplier.•Reconstitute the PK—Add the required volume of PK Buffer according to the recommendations of the supplier.Perform the purification procedurebine the following components in the order indicated, then immediately proceed to the nextstep.b.Vortex for 15 seconds.c.Incubate at 70℃ for 30 minutes.d.Allow the tubes to cool, then centrifuge the samples briefly to bring down condensation.e.Add 200 μL of B3 Buffer, then vortex for 15 seconds.f.Incubate at 70℃ for 10 minutes.g.Allow the tubes to cool, then centrifuge briefly.h.Add 200 μL of 96–100% ethanol to each sample, vortex for 15 seconds, then briefly centrifugeto collect the contents.1Lyse, then homogenize the samplesa.Insert a NucleoSpin ™Tissue kit column into a collection tube, then transfer the entire samplevolume to the column.b.Cap the column, then centrifuge the assembly at 11,000 × g for 1 minute.c.Discard the collection tube, then place the column on a new collection tube.2Bind the DNA to the columna.Add 500 μL of BW Buffer to each column, cap the column, then centrifuge at 11,000 × g for1 minute.b.Discard the collection tube, then place the column on a new collection tube.c.Add 500 μL of B5 Buffer to each column, cap the column, then centrifuge at 11,000 × g for1 minute d.Discard the collection tube, then place the column on a new collection tube.e.Centrifuge at 11,000 × g for 3 minutes to dry the membrane.f.Discard the collection tube.g.Place the column on a new 1.5‑mL microtube, then add 200 μL of BE Buffer.3Wash, then elute the DNA3Wash, then elute the DNA (continued)h.Cap the column, then incubate at room temperature for 1 minute.i.Centrifuge at 6,000 × g for 1 minute, then discard the column.The purified DNA is in the microtube.Store the purified DNA at 2–8°C for immediate use or below –16°C for long-term storage.Good laboratory practices for PCR and RT-PCR•Wear clean gloves and a clean lab coat.–Do not wear the same gloves and lab coat that you have previously used when handling amplified products or preparing samples.•Change gloves if you suspect that they are contaminated.•Maintain separate areas and dedicated equipment and supplies for:–Sample preparation and reaction setup.–Amplification and analysis of products.•Do not bring amplified products into the reaction setup area.•Open and close all sample tubes carefully. Avoid splashing or spraying samples.•Keep reactions and components capped as much as possible.•Use a positive-displacement pipettor or aerosol‑resistant barrier pipette tips.•Clean lab benches and equipment periodically with 10% bleach solution or DNA decontamination solution.Appendix A Purification with the KingFisher™ Duo Prime or KingFisher™ mL instrumentFollow this procedure for purification with the MagMAX™ CORE Nucleic Acid Purification Kit, using the KingFisher™ Duo Prime or KingFisher™ mL instrument.Required materials not suppliedTable 9 Materials required for processing on the KingFisher™ Duo Prime and KingFisher™ mL instruments[1]Unless otherwise indicated, all materials are available through . "MLS" indicates that the material is available from or another major laboratorysupplier.[2]Included in the KingFisher™ Duo Combi pack (Cat. No. 97003530).Purification procedureNote: When performing this procedure for processing on the KingFisher™ mL instrument, mix samples by pipetting up and down. Do not use a plate shaker with the large tube strips required by this instrument.1.Follow the protocol, starting with sample lysate preparation through combining the samples with beads and lysis solution.Note: Do not set up processing plates or tubes before preparing samples.2.Add MagMAX™ CORE Wash Solutions and MagMAX™ CORE Elution Buffer to the indicated positions, according to your instrument.Load the Tip Comb and all of the plates or tube strips at the same time. The instrument does not prompt you to load itemsindividually.Table 10 Plate setup: KingFisher™ Duo Prime instrument[1]Ensure that the elution strip is placed in the correct direction in the elution block.[2]Placed on the heating element.Table 11 Tube strip setup: KingFisher™ mL instrument3.Select the appropriate script on the instrument (see “Download and install the script” on page 6).4.Start the run, then load the prepared plates or tube strips in the appropriate positions when prompted by the instrument.Store purified nucleic acid on ice for immediate use, at −20°C for up to 1 month, or at −80°C for long‑term storage.Appendix B Documentation and supportCustomer and technical supportVisit /support for the latest service and support information.•Worldwide contact telephone numbers•Product support information–Product FAQs–Software, patches, and updates–Training for many applications and instruments•Order and web support•Product documentation–User guides, manuals, and protocols–Certificates of Analysis–Safety Data Sheets (SDSs; also known as MSDSs)Note: For SDSs for reagents and chemicals from other manufacturers, contact the manufacturer.Limited product warrantyLife Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies' General Terms and Conditions of Sale at /us/en/home/global/terms-and-conditions.html. If you have any questions, please contact Life Technologies at /support.Corporate entity: Life Technologies Corporation | Carlsbad, CA 92008 USA | Toll Free in USA 1 800 955 6288The information in this guide is subject to change without notice.DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.Revision history: Pub. No. MAN0019166Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these products, you accept the terms and conditions of all applicable Limited Use Label Licenses.©2020 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. FastPrep‑24 is a trademark of MP Biomedicals, LLC. Precellys is a trademark of Bertin Technologies. QIAamp is a trademark of QIAGEN GmbH. Nucleospin is a trademark of MACHEREY‑NAGEL./support | /askaquestion。

ColiComplete 产品说明书

Page 1 of 2 ColiComplete ®AOAC Official Method 992.30General DescriptionColiComplete ® contains 5-bromo-4-chloro-3-indolyl-ß-Dgalactopyranoside (X-Gal) and 4-methyl umbelliferyl-ß-D-glucuronide (MUG). Discs are added to LST inoculated with selected dilutions of samples. Samples are incubated at 35–37 °C and examined after 24 and 48 ±2 h for confirmed total coliforms and after 30 ±2 h for confirmed E. coli results. ß-Galactosidase, from coliforms present in samples, cleaves X-Gal into 5-bromo-4-chloro-indoxyl intermediate which undergoes oxidation to yield water-insoluble blue dimer, visually detectable on disc or in surrounding medium as confirmed positive result for total coliform activity. ß-Glucuronidase, from E. coli present in samples, cleaves MUG into glucuronide and methyl umbelliferone which fluoresces under long wave UV light (366 nm) as confirmed positive result for E. coli presence.NOTE : As E. coli O157:H7 does not produce ß-glucuronidase, ColiComplete ® is not suitable for the detection of E. coli O157:H7.A. Sample PreparationPrepare appropriate serial dilutions as indicated in FDA Bacteriological Analytical Manual (BAM), or AOAC Official Methods of Analysis according to sample type.B. InoculationInoculate LST tubes with appropriate sample dilution series selected to determine MPN levels or presence/absence of total coliforms and E. coli in sample. Aseptically add a single ColiComplete ® disc to each tube. Incubate at 35–37 °C.C. Reading ColiComplete ®a. For total coliforms — After at least 24 h incubation, examine each tube for visually detectable blue color on disc or in surrounding medium. Presence of blue color indicates confirmed positive result for total coliforms.NOTE: A wide range of blue color intensity may be expected, depending on sample composition and microflora. All blue reactions are positive regardless of intensity of color.Reincubate at 35–37 °C. After additional 24 ±2 h re-examine. Continued absence of blue indicates negative result; presence of blue indicates confirmed positive result for total coliforms. Read and record the MPN code or presence/absence of total coliforms in the sample.b. For E.coli — After 30 ±2 h from start of initial incubation, examine tubes under long-wave UV light (366 nm). Fluorescent tubes indicate confirmed positive result for E. coli. Read and record the MPN code or presence/absence of E. coli in the sample.D. CONTROLSPositive and negative controls should be used to facilitate interpretation of MUG fluorescent reaction. Use one known positive E. coli tube and two negative controls - one non -E. coli /coliform tube (e.g., Klebsiella spp.) and one uninoculated media tube.NOTE: Use borosilicate glass tubes, flint glass gives fluorescence that may be misinterpreted for a positive result.Lit. No. MK_UG4655EN Merck KGaAFrankfurter Strasse 25064293 DarmstadtGermanyPage 2 of 2 E. Method Modification for Certain JuicesApplicable to juice products/processors which rely on treatments that do not come into direct contact with all parts of the juice, as contained in 21 CFR Part 120: Rules and Regulations. Hazard Analysis and Critical Control Point (HAACP); Procedures for the Safe and Sanitary Processing and Importing of Juice; Final Rule. Vol 66 No. 13. 6137-6202. Use the modified method “Analysis for Escherichia coli in Citrus Juices - Modifi cation of AOAC Official Method 992.30” as stated in Section 120.25 (a).F. StorageStore unused discs at 2–8 °C (36–46 °F) in a sealed container, with desiccant.G. DisposalAfter use, all tubes must be steam-sterilized at 121 °C for at least 30 min before discarding. For in-vitro diagnostic use only.Manufacturing EntityBioControl Systems, Inc, 12822 SE 32nd St, Bellevue, WA 98005, USA.BioControl Systems, Inc is an affiliate of Merck KGaA, Darmstadt, Germany.。

ARIES ARIES-P -Ver.04- 8 0 2 7 9 0 8 1 1 3 7 4 0 产

ISTRUZIONI D'USO E DI INSTALLAZIONE INSTALLATION AND USER'S MANUALINSTRUCTIONS D'UTILISATION ET D'INSTALLATION INSTALLATIONS-UND GEBRAUCHSANLEITUNG INSTRUCCIONES DE USO Y DE INSTALACION INSTRUÇÕES DE USO E DE INSTALAÇÃOCENTRALINA DI COMANDO D811184A ver. 04 08-02-02I CONTROL UNIT GB UNITÉ DE COMMANDE F STEUERZENTRALE D CENTRAL DE MANDO E CENTRAL DO MANDOP ARIES - ARIES P8027908113740a“WARNINGS” leaflet and an “INSTRUCTION MANUAL”.These should both be read carefully as they provide important information about safety, installation, operation and maintenance. This product complies with the recognised technical standards and safety regulations. We declare that this product is in conformity with the following European Directives: 89/336/EEC and 73/23/EEC (and subsequent amendments).1) GENERAL OUTLINEThe ARIES control unit has been designed for swing gates. It can be used for one or two gate controllers.The control unit mod. ARIES P can also be used to perform opening of a single actuator while keeping the other one closed (pedestrian access).2) FUNCTIONSSTOP: In all cases: it stops the gate until a new start command is given.PHOT:Functions can be set with Dip-Switch.Activated during closing.Activated during opening and closing.Rapid closingON: When the position of the gate photocells is exceeded, during both opening and closing, the gate automatically starts to close even if TCA is activated. We recommend setting DIP3 to ON (photocells only activated during closing).Blocks impulsesON: During opening, START commands are not accepted.OFF: During opening, START commands are accepted.PhotocellsON: Photocells only activated during closing.OFF: Photocells activated during opening and closing.Automatic closing time (TCA)ON: Automatic closing activated (can be adjusted from 0 to 90s)Preallarm (mod. ARIES P only)ON: The flashing light turns on abt 3 seconds before the motors start.FOR THE INSTALLER: check the boxes you are interested in.START:four-step logic Gate closedGate openDuring openingDuring closingAfter stop START: two-step logic SCA: Gate open indicating lightit opens it opensit stops and activates TCAit closesit stops and does not activate TCAit starts opening it stops and activats TCA (if activated)it closesit opensit opensoffononflashingATTENTION:Dip non used in mod. ARIES (always in OFF set).3) MAINTENANCE AND DEMOLITIONThe maintenance of the system should only be carried out by qualified personnel regularly. The materials making up the set and its packing must be disposed of according to the regulations in force.Batteries must be properly disposed of.WARNINGSCorrect controller operation is only ensured when the data contained in the present manual are observed. The company is not to be held responsible for any damage resulting from failure to observe the installation standards and the instructions contained in the present manual.The descriptions and illustrations contained in the present manual are not binding. The Company reserves the right to make any alterations deemed appropriate for the technical, manufacturing and commercial improvement of the product, while leaving the essential product features unchanged, at any time and without undertaking to update the present publication.D 811184A _04Thank you for buying this product, our company is sure that you will be more than satisfied with the product ’s performance. The product is supplied with a “WARNINGS ” leaflet and an “INSTRUCTION MANUAL ”.These should both be read carefully as they provide important information about safety, installation, operation and maintenance.This product complies with the recognised technical standards and safety regulations. We declare that this product is in conformity with the following European Directives: 89/336/EEC and 73/23/EEC (and subsequent amendments).1) GENERAL OUTLINEThe ARIES control unit has been designed for swing gates. It can be used for one or two gate controllers.The control unit mod. ARIES P can also be used to perform opening of a single actuator while keeping the other one closed (pedestrian access).2) GENERAL SAFETYWARNING! An incorrect installation or improper use of the product can cause damage to persons, animals or things.•The “Warnings ” leaflet and “Instruction booklet ” supplied with this product should be read carefully as they provide important information about safety, installation, use and maintenance.•Scrap packing materials (plastic, cardboard, polystyrene etc) according to the provisions set out by current standards. Keep nylon or polystyrene bags out of children ’s reach.•Keep the instructions together with the technical brochure for future reference.•This product was exclusively designed and manufactured for the use specified in the present documentation. Any other use not specified in this documentation could damage the product and be dangerous.•The Company declines all responsibility for any consequences resulting from improper use of the product, or use which is different from that expected and specified in the present documentation.•Do not install the product in explosive atmosphere.•The Company declines all responsibility for any consequences resulting from failure to observe Good Technical Practice when constructing closing structures (door, gates etc.), as well as from any deformation which might occur during use.•The installation must comply with the provisions set out by the following European Directives: 89/336/EEC, 73/23/EEC, 98/37/ECC and subsequent amendments.•Disconnect the electrical power supply before carrying out any work on the installation. Also disconnect any buffer batteries, if fitted.•Fit an omnipolar or magnetothermal switch on the mains power supply,having a contact opening distance equal to or greater than 3mm.•Check that a differential switch with a 0.03A threshold is fitted just before the power supply mains.•Check that earthing is carried out correctly: connect all metal parts for closure (doors, gates etc.) and all system components provided with an earth terminal.•The Company declines all responsibility with respect to the automation safety and correct operation when other manufacturers ’ components are used.•Only use original parts for any maintenance or repair operation.•Do not modify the automation components, unless explicitly authorised by the company.•Instruct the product user about the control systems provided and the manual opening operation in case of emergency.•Do not allow persons or children to remain in the automation operation area.•Keep radio control or other control devices out of children ’s reach, in order to avoid unintentional automation activation.•The user must avoid any attempt to carry out work or repair on the automation system, and always request the assistance of qualified personnel.•Anything which is not expressly provided for in the present instructions,is not allowed.3) TECHNICAL SPECIFICATIONSPower supply:...............................................................230V ±10% 50Hz Absorption on empty:.................................................................0.5A max Output power for accessories:..........................................24V~ 6VA max Max relay current:................................................................................8A Max power of motors:...............................................................300 W x 2Torque limiter:.................................................Self-transformer with 4 pos Limit switch:................................................................Adjustable run timePanel dimensions:.........................................................................See fig.1Cabinet protection:............................................................................IP55Working temperature:...............................................................-20 +55°C 4) TERMINAL BOARD CONNECTIONS(Fig.2)CAUTION: Keep the low voltage connections completely separated from the power supply connections.Fig.3 shows the fixing and connection method of the drive condensers whenever they are not fitted to the motor.JP51-2 Single-phase power supply 230V ±10%, 50 Hz (1=L/2=N).For connection to the mains use a multiple-pole cable with a minimum cross section of 3x1.5mm 2 of the type indicated in the above-mentioned standard (by way of example, if the cable is not shielded it must be at least equivalent to H07 RN-F while, if shielded, it must be at least equivalent to H05 VV-F with a cross section of 3x1.5mm 2).JP33-4 (mod.ARIES-P) 230V 40W max. blinker connection.5-6 (mod.ARIES) 230V 40W max. blinker connection.7-8-9 Motor M1 connection - 8 common, 7-9 start.10-11-12 Motor M2(r) connection - 11 common, 10-12 start.JP413-14 Open-close button and key switch (N.O.).13-15 Stop button (N.C.). If unused, leave bridged.13-16 Photocell or pneumatic edge input (N.C.). If unused, leave bridged.17-18 24V 3W max. gate open warning light.18-19 24V~ 0.25A max. (6VA) output (for supplying photocell or other device).20-21 Antenna input for radio-receiver board (20 signal - 21 braid).22 Common terminal (equivalent to terminal 13).23 Terminal for pedestrian control. It moves the leaf of motor M2 connected to terminal 10-11-12. This terminal is available only in ARIES-P control unit.JP225-26 2nd radio channel output of the double-channel receiver board (terminals not fitted on ARIES but fitted on ARIES-P) contact N.O.JP1 Radio-receiver board connector 1-2 channels.5) FUNCTIONSDL1:Power-on LedIt is switched on when the board is electrically powered.START: four-step logic: (DIP5 OFF)gate closed:..................................................................................it opens during opening:............................................... it stops and activates TCA gate open:................................................................................... it closes during closing:.................................... it stops and does not activate TCA after stop:.........................................................................it starts opening START: two-step logic: (DIP5 ON)gate closed:..................................................................................it opens during opening:................................it stops and activats TCA (if activated)gate open:....................................................................................it closes during closing:..............................................................................it opens after stop:.....................................................................................it opens STOP: In all cases: it stops the gate until a new start command is given.PHOT:Functions can be set with DIP-SWITCH.Activated during closing if DIP3-ON.Activated during opening and closing if DIP3-OFF.SCA: Gate open indicating light.with gate closed:...................................................................................off when gate is opening:...........................................................................on with gate open:.......................................................................................on when gate is closing:.....................................................................flashing 6) DIP-SWITCH SELECTION DIP1 Rapid closingON: When the position of the gate photocells is exceeded, during both opening and closing, the gate automatically starts to close even if TCA is activated. We recommend setting DIP3 to ON (photocells only activated during closing).OFF: Function not activated.DIP2 Blocks impulsesON: During opening, START commands are not accepted.OFF: During opening, START commands are accepted.DIP3 PhotocellsON: Photocells only activated during closing.OFF: Photocells activated during opening and closing.D 811184A _04DIP4 Automatic closing time (TCA)ON: Automatic closing activated (can be adjusted from 0 to 90s).OFF: Automatic closing not activated.DIP5 Control logicON: 2-step logic is activated (see start paragraph).OFF: 4-step logic is activated (see start paragraph).DIP6: Preallarm (mod.ARIES P only)ON: The flashing light turns on abt 3 seconds before the motors start.OFF The flashing light turns on simultaneously with the start of the motors.ATTENTION:Dip non used in mod. ARIES (always in OFF set).7) TRIMMER ADJUSTMENTTCA This adjusts the automatic closing time, after which time the gate automatically closes (can be adjusted from 0 to 90s).TW This adjusts the motor working time, after which time the motor stops (can be adjusted from 0 to 40s).TDELAY This adjusts the closing delay time of the second motor (M2).8) MOTOR TORQUE ADJUSTMENTThe ARIES control unit has electric torque adjustment which allows the motor force to be adjusted.The adjustment should be set for the minimum force required to carry out the opening and closing strokes completely.Adjustment is carried out by moving the connection 55 (fig.3) on the tran-sformer sockets as described below:Pos.T1 1st TORQUE (MINIMUM TORQUE)Pos.T2 2nd TORQUE Pos.T3 3rd TORQUEPos.T4 4th TORQUE (MAXIMUM TORQUE)4 motor torque values can be obtained.To gain access to the torque adjustment sockets, disconnect the mains supply and remove the protective case “P ” of the transfomer.CAUTION: Excessive torque adjustment may jeopardise the anti-squash safety function. On the other hand insufficient torque adjustment may not guarantee correct opening or closing strokes.9) MAINTENANCE AND DEMOLITIONThe maintenance of the system should only be carried out by qualified personnel regularly. The materials making up the set and its packing must be disposed of according to the regulations in force.Batteries must be properly disposed of.WARNINGSCorrect controller operation is only ensured when the data contained in the present manual are observed. The company is not to be held responsible for any damage resulting from failure to observe the installation standards and the instructions contained in the present manual.The descriptions and illustrations contained in the present manual are not binding. The Company reserves the right to make any alterations deemed appropriate for the technical, manufacturing and commercial improvement of the product, while leaving the essential product features unchanged, at any time and without undertaking to update the present publication.D811184A_04ARIES/ARIES-P - Ver. 04 -23。

EXFO 2000 系列以太网测试仪中文说明书

S P E C S H E ETKEY FEATURES AND BENEFITSAccelerate Ethernet service activation with bidirectional EtherSAM (Y .156) and RFC 2544 test suites, multistream trafﬁ c generation, Through mode and bit-error-rate (BER) testing Experience unprecedented conﬁ guration simplicity with hybrid touchscreen/keypad navigation and data entry Increase technician autonomy and productivity with intelligent discovery of remote EXFO Ethernet testers, as well as in-service testing via dual-port Through mode Eliminate errors in data interpretation with revolutionary new GUI on 7-inch TFT screen, historical event logger, visual gauges and 3D-icon depictions of pass/fail outcomesSimplify reporting with integrated Wi-Fi and Bluetooth connectivity capabilitiesIntegrated applications to test VoIP services, and additional IP test utilities including VLAN scan and LAN discovery via EXpert VoIP and EXpert IP test toolsSupport for packet capture and analysis, wireless troubleshooting and TCP throughput testingExtend ﬁ eld testing operations with compact, lightweight platform equipped with long-duration battery packFTB-860 NetBlazer Series Ethernet TestersPOWERFUL, FAST, INTUITIVE ETHERNET TESTINGeld technicians comprehensive, yet simple test suites to quickly and easily turn up, validate and troubleshoot Ethernet services, with full EtherSAM capabilities, from 10 Mbit/s to 10 Gbit/s.EXFO FTB-1 FTB-860G SpecsProvided by THE ULTRA-PORTABLE CHOICE FOR HIGH-SPEED ETHERNET TESTINGThe ongoing deployment of GigE and 10 GigE circuits across access and metro networks demands a testing solution that seamlessly adapts to either operating environment—without sacrificing portability, speed or cost—in order to guarantee the performance and quality of service (QoS) metrics of these services.Leveraging the powerful, intelligent FTB-1 handheld platform, the NetBlazer series streamlines processes and empowers field technicians to seamlessly transition between 10/100/1000/10000 interfaces to rapidly adapt to a variety of networking environments.Powerful and FastThe NetBlazer series is a portfolio of fully integrated 10 Mbit/s to 10 Gbit/s handheld Ethernet testers. Available in three hardware configurations, each FTB-860x offers the industry’s largest TFT screen with unprecedented configuration simplicity via hybrid touchscreen/keypad navigation. Platform connectivity is abundant via Wi-Fi, Bluetooth, Gigabit Ethernet or USB ports, making it accessible in any environment.FTB-860G: 10 M BIT /S TO 10 G BIT/SIf the need is for full Ethernet coverage from 10 Mbit/s up to 10 Gbit/s, › 10 Base-T to 10 gigabit testing› IPv6 testingFTB-860: GIGABIT ETHERNETIf the need is purely for Gigabit Ethernet coverage, then the FTB-860 is › 10 Base-T to 1 gigabit testing› IPv6 testingFTB-860GL: 10 M BIT/S TO 10 G BIT/S LOOPBACK ONLYCombined with the FTB-860G or FTB-860, the FTB-860GL is the most cost-effective solution for GigE and 10 GigE intelligent loopback testing; it supports bidirectional EtherSAM and RFC 2544 testing and offers five › 10 Base-T to 10 gigabit loopback› EtherSAM (bidirectional partner)*› RFC 2544 (bidirectional partner)› Intelligent autodiscovery› IPv6 testing› Ping/traceroute* Contact your EXFO representative to confirm availability.Setting a New GUI Standard: Unprecedented Simplicity in Configuration Setup and NavigationIntelligent Situational Configuration Setup›G uides technicians through complete, accurate testingprocesses(suggestion prompts, help guides, etc.)›R educes navigation by combining associated testingfunctions on a single screen›I ntelligent autodiscovery allows a single technician to performend-to-end testingDedicated Quick-Action Buttons›Remote discovery to ﬁ nd all the other EXFO units›Laser on/off›T est reset to clear the results and statistics while running a test ›Report generation›Save or load test conﬁ gurations›Quick error injectionAssorted Notifications›Clear indication of link status for single or dual ports›Negotiated speed display for single or dual ports›O ptical power status available at all times for single or dual ports›Pass/fail indication at all times for all testsStreamlined Navigation›R emote discovery button available at all times; no reason to leave your current location to scan for a remote unit›T esting status can be maximized to ﬁ ll the entire screen by simply clicking on the alarm status button; whether the unit is in your hand or across the room, test results can be easily determined with a simple glance at the display screen›R FC 2544 conﬁguration is maximized in a single page;no need to navigate through multiple screens to conﬁindividual subtests›R FC 2544 results and graphs are also maximized in a single page; no need to navigate through multiple screens to viewindividual RFC subtest results FO unitswhile running a testdual portsal portstimes for single mes; no reason toemote unite entire screen by ; whether the unit sults can be easily splay screenn a single page; eens to conﬁ gure ximized in a single e screens to viewRAPID, ACCURATE TEST RESULTS AT YOUR FINGERTIPSKey FeaturesIntelligent Network Discovery ModeUsing any NetBlazer series test set, you can single-handedly scan the network and connect to any available EXFO datacom remote tester. Simply select the unit to be tested and choose whether you want traffic to be looped back via Smart Loopback or Dual Test Set for simultaneous bidirectional EtherSAM and RFC 2544 results. No more need for an additional technician at the far end to relay critical information—the NetBlazer products take care of it all.Smart Loopback FlexibilityThe Smart Loopback functionality has been enhanced to offer five distinct loopback modes. Whether you are looking to pinpoint loopback traffic from a UDP or TCP layer, or all the way down to a completely promiscuous mode (Transparent Loopback mode), NetBlazer has the flexibility to adjust for all unique loopback situations.Global Pass/Fail AnalysisThe NetBlazer series provides real-time pass/fail status via text or icons. Clicking on the pass/fail indicator maximizes this important status to full screen, providing instant, easily understood notification whether the unit is in the technician’s hands or across the room.Remembering the Last IP or MAC AddressesField technicians have enough things to worry about and don’t always have the luxury of time to enter the same IP or MAC address test after test. The NetBlazer series remembers the last 10 MAC, IPv4 and IPv6 addresses as well as J0/J1 traces for 10G WAN, even afterthe unit has been rebooted.Traffic GenerationUnparalleled analog visual gauges combined with user-defined thresholds show instantaneously whether or not the test traffic is in or out of expected ranges.Additionally, bandwidth and frame size can be modified on-the-fly without navigating away to a different page, giving technicians instantaneous reaction on the gauges. Traffic generation brings together over 10 critical stats in a very visual and organizedfashion, ensuring that technicians can quickly and easily interpret the outcome of the test.The analog gauges are lined with Green and Red layers to represent the expected thresholds.Real-time bandwidth and frame-size adjustment.Overall pass/fail assessment.Throughput, jitter and latency with visual pass/fail thresholds,analog gauges and digitalreadouts.Frame loss and out-of-sequence notification.Multistream ConfigurationConfiguring multiple streams with proper COS and QOS bits can be a complex task. NetBlazer makes it simpler, with all streams easily selectable and configurable from one location. With large icons located throughout the stream pages, configuration becomes as simple as a touch of a finger. Technicians can define one configuration profile and apply it to all the background streams simultaneously. From there, it is just a matter of making slight tweaks as needed rather than complete configuration profiles per stream.Through ModeThrough mode testing consists of passing traffic through either of the NetBlazer’s two 100/1000 Base-X ports or the two 10/100/1000 Base-T ports for in-service troubleshooting of live traffic between the carrier/service provider network and the customer network. Through mode allows technicians to access circuits under test without the need for a splitter.Supporting 10 Gigabit EthernetThe 10 G igabit Ethernet interface is available in both 10 G igE LAN and 10 G igE WAN modes via a single SFP+ transceiver. All Ethernet testing applications—from BER testing to the full EtherSAM suite—are available for both IPv4 and IPv6. Unique to the 10 GigE WAN interface is the ability to send and monitor SONET/SDH J0/J1 traces and the path signal label (C2). The WAN interface can also monitor SONET and SDH alarms and errors.E THER SAM: THE NEW STANDARD IN ETHERNET TESTINGUntil now, RFC 2544 has been the most widely used Ethernet testing methodology. However it was designed for network device testing in the lab, not for service testing in the field. ITU-T Y.156sam is the newly introduced draft standard for turning up and troubleshooting carrier Ethernet services. It has a number of advantages over RFC 2544, including validation of critical SLA criteria such as packet jitter and QoS measurements. This methodology is also significantly faster, therefore saving time and resources while optimizing QoS.EXFO’s EtherSAM test suite—based on the draft ITU-T Y.156sam Ethernet service activation methodology—provides comprehensive field testing for mobile backhaul and commercial services.Contrary to other methodologies, EtherSAM supports new multiservice offerings. It can simulate all types of services that will run on the network and simultaneously qualify all key SLA parameters for each of these services. Moreover, it validates the QoS mechanisms provisioned in the network to prioritize the different service types, resulting in better troubleshooting, more accurate validation and much faster deployment. EtherSAM is comprised of two phases, the network configuration test and the service test.Network Configuration TestThe network configuration test consists of sequentially testing each service. It validates that the service is properly provisioned and that all specific KPIs or SLA parameters are met.Service TestOnce the configuration of each individual service is validated, the service test simultaneously validates the quality of all the services over time.EtherSAM Bidirectional ResultsEXFO’s EtherSAM approach proves even more powerful as it executes the complete ITU-T Y.156sam test with bidirectional measurements. Key SLA parameters are measured independently in each test direction, thus providing 100 % first-time-rightservice activation—the highest level of confidence in service testing.EX PERT TEST TOOLSEXpert Test Tools is a series of platform-based software testing tools that enhance the value of the FTB-1 platform, providing additional testing capabilities without the need for additional modules or units.The EXpert VoIP Test Tools generates a voice-over-IP call directly from the test platform to validateperformance during service turn-up and troubleshooting.›Supports a wide range of signaling protocols, including SIP, SCCP, H.248/Megaco and H.323 ›Supports MOS and R-factor quality metrics› Simplifies testing with configurable pass/fail thresholds and RTP metricsThe EXpert IP Test Tools integrates six commonly used datacom test tools into one platform-based application to ensure field technicians are prepared for a wide-range of testing needs. › Rapidly perform debugging sequences with VLAN scan and LAN discovery› Validate end-to-end ping and traceroute› Verify FTP performance and HTTP availabilityTEST TOOLS IPEXpert TEST TOOLS VoIPOPTICAL INTERFACESTwo ports: 100M and GigEAvailable wavelengths (nm)850, 1310 and 1550100 Base-FX100 Base-LX1000 Base-SX1000 Base-LX1000 Base-ZX1000 Base-BX10-D1000 Base-BX10-USFP+ OPTICAL INTERFACES (10G)10G Base-SR/SW10G Base-LR/LW 10G Base-ER/EW Wavelength (nm)85013101550Tx level (dBm)–5 to –1–8 to 0.5–4.7 to 4.0SPECIFICATIONSELECTRICAL INTERFACESTwo ports: 10/100 Base-T half/full duplex, 1000 Base-T full duplexGENERAL SPECIFICATIONSSize (H x W x D)130 mm x 36 mm x 252 mm (5 1/8 in x 1 7/16 in x 9 15/16 in)Weight (with battery) 0.58 kg (1.3 lb)TemperatureTESTINGEtherSAM (Y.156sam)Network configuration and service test as per ITU-T Y.156sam. Tests can be performed using remote loopback orADDITIONAL FEATURESOptical power measurement Supports optical power measurement at all times; displayed in dBm.UPGRADESFTB-8590SFP modules GigE/FC/2FC at 850 nm, MM, <500 mEXFO is certified ISO 9001 and attests to the quality of these products. This device complies with Part 15 of the FCC Rules. Operation is subject to the following two conditions: (1) this device may not cause harmful interference, and (2) this device must accept any interference received, including interference that may cause undesired operation. EXFO has made every effort to ensure that the information contained in this specification sheet is accurate. However, we accept no responsibility for any errors or omissions, and we reserve the right to modify design, characteristics and products at any time without obligation. Units of measurement in this document conform to SI standards and practices. In addition, all of EXFO’s manufactured products are compliant with the European Union’s WEEE directive. For more information, please visit /recycle. Contact EXFO for prices and availability or to obtain the phone number of your local EXFO distributor. For the most recent version of this spec sheet, please go to the EXFO website at /specs .In case of discrepancy, the Web version takes precedence over any printed literature.EXFO Corporate Headquarters > 400 Godin Avenue, Quebec City (Quebec) G1M 2K2 CANADA | Tel.: +1 418 683-0211 | Fax: +1 418 683-2170 |*************Toll-free: +1 800 663-3936 (USA and Canada) | EXFO America 3701 Plano Parkway, Suite 160Plano, TX 75075 USA Tel.: +1 800 663-3936 Fax: +1 972 836-0164 EXFO Asia 151 Chin Swee Road, #03-29 Manhattan House SINGAPORE 169876Tel.: +65 6333 8241 Fax: +65 6333 8242EXFO China 36 North, 3rd Ring Road East, Dongcheng District Beijing 100013 P. R. CHINATel.: + 86 10 5825 7755 Fax: +86 10 5825 7722Room 1207, Tower C, Global Trade Center EXFO Europe Omega Enterprise Park, Electron Way Chandlers Ford, Hampshire S053 4SE ENGLAND Tel.: +44 2380 246810 Fax: +44 2380 246801EXFO NetHawkElektroniikkatie 2 FI-90590 Oulu, FINLAND Tel.: +358 (0)403 010 300 Fax: +358 (0)8 564 5203EXFO Service Assurance270 Billerica RoadChelmsford, MA 01824 USATel.: +1 978 367-5600Fax: +1 978 367-5700FTB-860 NetBlazer Series Ethernet TestersORDERING INFORMATIONSPFTB860Series.1AN© 2010 EXFO Inc. All rights reserved.Printed in Canada 10/09FTB-860G-XX -XX -XXNotesa. Requires purchase of SFP.b. Requires purchase of SFP+.。

提示工程(prompt)

客户问题：你好，我有一个关于我的汽车保险的问题。分类主题：1
客户问题：{在此插入新问题} 分类主题：
之前
单个家庭的年用水需求是多少？包含四个在家的人的家庭每年平均200天，使用量平均为100 每天多少升水？
单户家庭的年用水需求包含四个人，他们平均在家的时间为每年200天，平均使用100升的每天的用水量为每年800万升。
2nd prompt—答案提取
一个普通人在一分钟内能打出25拳。一场比赛持续5个回合，每回合3分钟。有多少他投掷了多少拳？
让我们一步一步地思考并解释这个计算过程。一步一步地。
在一分钟内，乔打了25拳。
在三分钟内，乔打了3 * 25 = 75拳。
在五轮中，乔投掷了5*75 = 375次拳。
在一分钟内，乔打了25拳。在三分钟内，乔打了3 * 25 = 75拳。在五轮中，乔投掷了5*75 = 375次拳。
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摘要
请总结信息关于产品 [{'text': 'iPhone', 'type': '产品'}], {'text': '苹果', 'type': 'ORGANIZATION’}]
iPhone是一系列智能手机。由苹果设计并销售 Inc. 这些设备结合了计算机、相机和互联网单一设备中的连接性，以及以其简洁的设计而闻名用户友好的界面。
人工智能机器学习深度学习
生成式人工智能
1956
人工智能
计算机科学领域中致力于创造的分支能够复制或超越人类智能的智能机器人类智能
1997
机器学习
人工智能的子集，使机器能够从现有数据，并在该数据的基础上进行改进，以使其决策或预测

Sysmex XN系列和SP-10维护指南说明书

Sysmex XN Series and SP-10 MaintenanceDocument Number: RHEAHS14010MUL Revision Number: 2.50 Document Type: Procedure Effective Date: 6/17/2022 2:26:04 PM Location: 3. APL Folder Structure\Hematology HE\CBC and Differential\Sysmex XN - 59\Edmonton ZoneRHEAHS14010MUL Sysmex XN Series and SP-10 MaintenanceAPPLICABILITYThis document is applicable to APL staff at the following Edmonton Zone sites: GNH,RAH, SGH and UAH.PURPOSEThis procedure describes how to perform the daily clean, shutdown, startup and maintenance (daily, weekly and monthly) of the Sysmex XN series and SP-10 analyzers.MATERIALSPROCEDURE4.1. XN Series MaintenanceAll maintenance procedures must be signed off on the Sysmex XN Series Maintenance form.4.1.1. Daily4.1.2. Weekly4.1.3. Monthly4.1.4. Every Six Months4.1.5. Yearly or As Needed4.2. SP-10 Maintenance (RAH and UAH only)All maintenance procedures should be signed off on the Sysmex SP-10 Maintenance form.4.2.1. DailyWhen in shutdown, remove the trap chamber from the right side of4.2.2.Weekly4.2.3.As NeededREFERENCES1. Sysmex XN-1000 / XN-2000 Automated Hematology Analyzers CLSI Procedure, Document number:1004-LSS. Rev. 1, March 20132. XN Series (For XN-1000 system) Instructions for Use (North American Edition), November 2012.3. SP-10 Instructions for use.4. XN-9000 Instructions for use.RELATED DOCUMENTSRHEAHF14010MULA Sysmex XN Series MaintenanceRHEAHS14006MUL Sysmex XN Series - Performing QCRHEAHS14019MUL Sysmex XN Series - Manual Shutdown and StartupRHERHS00004MUL Inter-Instrument Variability for Hematology Coagulation and Special Coagulation AnalyzersRHERHF00002MUL Hematology Inter-Instrument Variability Excel WorksheetRQMPCS14000MUL Annual Autovalidation CheckRHEAHF14010MULB Sysmex SP-10 MaintenanceHE24-097 Evaluating Quality Control using the Beyond Care Quality Monitoring BCQM System。

Indradrive 系列故障代码

Error MessagesF9001 Error internal function call.F9002 Error internal RTOS function callF9003 WatchdogF9004 Hardware trapF8000 Fatal hardware errorF8010 Autom. commutation: Max. motion range when moving back F8011 Commutation offset could not be determinedF8012 Autom. commutation: Max. motion rangeF8013 Automatic commutation: Current too lowF8014 Automatic commutation: OvercurrentF8015 Automatic commutation: TimeoutF8016 Automatic commutation: Iteration without resultF8017 Automatic commutation: Incorrect commutation adjustment F8018 Device overtemperature shutdownF8022 Enc. 1: Enc. signals incorr. (can be cleared in ph. 2) F8023 Error mechanical link of encoder or motor connectionF8025 Overvoltage in power sectionF8027 Safe torque off while drive enabledF8028 Overcurrent in power sectionF8030 Safe stop 1 while drive enabledF8042 Encoder 2 error: Signal amplitude incorrectF8057 Device overload shutdownF8060 Overcurrent in power sectionF8064 Interruption of motor phaseF8067 Synchronization PWM-Timer wrongF8069 +/-15Volt DC errorF8070 +24Volt DC errorF8076 Error in error angle loopF8078 Speed loop error.F8079 Velocity limit value exceededF8091 Power section defectiveF8100 Error when initializing the parameter handlingF8102 Error when initializing power sectionF8118 Invalid power section/firmware combinationF8120 Invalid control section/firmware combinationF8122 Control section defectiveF8129 Incorrect optional module firmwareF8130 Firmware of option 2 of safety technology defectiveF8133 Error when checking interrupting circuitsF8134 SBS: Fatal errorF8135 SMD: Velocity exceededF8140 Fatal CCD error.F8201 Safety command for basic initialization incorrectF8203 Safety technology configuration parameter invalidF8813 Connection error mains chokeF8830 Power section errorF8838 Overcurrent external braking resistorF7010 Safely-limited increment exceededF7011 Safely-monitored position, exceeded in pos. DirectionF7012 Safely-monitored position, exceeded in neg. DirectionF7013 Safely-limited speed exceededF7020 Safe maximum speed exceededF7021 Safely-limited position exceededF7030 Position window Safe stop 2 exceededF7031 Incorrect direction of motionF7040 Validation error parameterized - effective thresholdF7041 Actual position value validation errorF7042 Validation error of safe operation modeF7043 Error of output stage interlockF7050 Time for stopping process exceeded8.3.15 F7051 Safely-monitored deceleration exceeded (159)8.4 Travel Range Errors (F6xxx) (161)8.4.1 Behavior in the Case of Travel Range Errors (161)8.4.2 F6010 PLC Runtime Error (162)8.4.3 F6024 Maximum braking time exceeded (163)8.4.4 F6028 Position limit value exceeded (overflow) (164)8.4.5 F6029 Positive position limit exceeded (164)8.4.6 F6030 Negative position limit exceeded (165)8.4.7 F6034 Emergency-Stop (166)8.4.8 F6042 Both travel range limit switches activated (167)8.4.9 F6043 Positive travel range limit switch activated (167)8.4.10 F6044 Negative travel range limit switch activated (168)8.4.11 F6140 CCD slave error (emergency halt) (169)8.5 Interface Errors (F4xxx) (169)8.5.1 Behavior in the Case of Interface Errors (169)8.5.2 F4001 Sync telegram failure (170)8.5.3 F4002 RTD telegram failure (171)8.5.4 F4003 Invalid communication phase shutdown (172)8.5.5 F4004 Error during phase progression (172)8.5.6 F4005 Error during phase regression (173)8.5.7 F4006 Phase switching without ready signal (173)8.5.8 F4009 Bus failure (173)8.5.9 F4012 Incorrect I/O length (175)8.5.10 F4016 PLC double real-time channel failure (176)8.5.11 F4017 S-III: Incorrect sequence during phase switch (176)8.5.12 F4034 Emergency-Stop (177)8.5.13 F4140 CCD communication error (178)8.6 Non-Fatal Safety Technology Errors (F3xxx) (178)8.6.1 Behavior in the Case of Non-Fatal Safety Technology Errors (178)8.6.2 F3111 Refer. missing when selecting safety related end pos (179)8.6.3 F3112 Safe reference missing (179)8.6.4 F3115 Brake check time interval exceeded (181)Troubleshooting Guide | Rexroth IndraDrive Electric Drivesand ControlsI Bosch Rexroth AG VII/XXIITable of ContentsPage8.6.5 F3116 Nominal load torque of holding system exceeded (182)8.6.6 F3117 Actual position values validation error (182)8.6.7 F3122 SBS: System error (183)8.6.8 F3123 SBS: Brake check missing (184)8.6.9 F3130 Error when checking input signals (185)8.6.10 F3131 Error when checking acknowledgment signal (185)8.6.11 F3132 Error when checking diagnostic output signal (186)8.6.12 F3133 Error when checking interrupting circuits (187)8.6.13 F3134 Dynamization time interval incorrect (188)8.6.14 F3135 Dynamization pulse width incorrect (189)8.6.15 F3140 Safety parameters validation error (192)8.6.16 F3141 Selection validation error (192)8.6.17 F3142 Activation time of enabling control exceeded (193)8.6.18 F3143 Safety command for clearing errors incorrect (194)8.6.19 F3144 Incorrect safety configuration (195)8.6.20 F3145 Error when unlocking the safety door (196)8.6.21 F3146 System error channel 2 (197)8.6.22 F3147 System error channel 1 (198)8.6.23 F3150 Safety command for system start incorrect (199)8.6.24 F3151 Safety command for system halt incorrect (200)8.6.25 F3152 Incorrect backup of safety technology data (201)8.6.26 F3160 Communication error of safe communication (202)8.7 Non-Fatal Errors (F2xxx) (202)8.7.1 Behavior in the Case of Non-Fatal Errors (202)8.7.2 F2002 Encoder assignment not allowed for synchronization (203)8.7.3 F2003 Motion step skipped (203)8.7.4 F2004 Error in MotionProfile (204)8.7.5 F2005 Cam table invalid (205)8.7.6 F2006 MMC was removed (206)8.7.7 F2007 Switching to non-initialized operation mode (206)8.7.8 F2008 RL The motor type has changed (207)8.7.9 F2009 PL Load parameter default values (208)8.7.10 F2010 Error when initializing digital I/O (-> S-0-0423) (209)8.7.11 F2011 PLC - Error no. 1 (210)8.7.12 F2012 PLC - Error no. 2 (210)8.7.13 F2013 PLC - Error no. 3 (211)8.7.14 F2014 PLC - Error no. 4 (211)8.7.15 F2018 Device overtemperature shutdown (211)8.7.16 F2019 Motor overtemperature shutdown (212)8.7.17 F2021 Motor temperature monitor defective (213)8.7.18 F2022 Device temperature monitor defective (214)8.7.19 F2025 Drive not ready for control (214)8.7.20 F2026 Undervoltage in power section (215)8.7.21 F2027 Excessive oscillation in DC bus (216)8.7.22 F2028 Excessive deviation (216)8.7.23 F2031 Encoder 1 error: Signal amplitude incorrect (217)VIII/XXII Bosch Rexroth AG | Electric Drivesand ControlsRexroth IndraDrive | Troubleshooting GuideTable of ContentsPage8.7.24 F2032 Validation error during commutation fine adjustment (217)8.7.25 F2033 External power supply X10 error (218)8.7.26 F2036 Excessive position feedback difference (219)8.7.27 F2037 Excessive position command difference (220)8.7.28 F2039 Maximum acceleration exceeded (220)8.7.29 F2040 Device overtemperature 2 shutdown (221)8.7.30 F2042 Encoder 2: Encoder signals incorrect (222)8.7.31 F2043 Measuring encoder: Encoder signals incorrect (222)8.7.32 F2044 External power supply X15 error (223)8.7.33 F2048 Low battery voltage (224)8.7.34 F2050 Overflow of target position preset memory (225)8.7.35 F2051 No sequential block in target position preset memory (225)8.7.36 F2053 Incr. encoder emulator: Pulse frequency too high (226)8.7.37 F2054 Incr. encoder emulator: Hardware error (226)8.7.38 F2055 External power supply dig. I/O error (227)8.7.39 F2057 Target position out of travel range (227)8.7.40 F2058 Internal overflow by positioning input (228)8.7.41 F2059 Incorrect command value direction when positioning (229)8.7.42 F2063 Internal overflow master axis generator (230)8.7.43 F2064 Incorrect cmd value direction master axis generator (230)8.7.44 F2067 Synchronization to master communication incorrect (231)8.7.45 F2068 Brake error (231)8.7.46 F2069 Error when releasing the motor holding brake (232)8.7.47 F2074 Actual pos. value 1 outside absolute encoder window (232)8.7.48 F2075 Actual pos. value 2 outside absolute encoder window (233)8.7.49 F2076 Actual pos. value 3 outside absolute encoder window (234)8.7.50 F2077 Current measurement trim wrong (235)8.7.51 F2086 Error supply module (236)8.7.52 F2087 Module group communication error (236)8.7.53 F2100 Incorrect access to command value memory (237)8.7.54 F2101 It was impossible to address MMC (237)8.7.55 F2102 It was impossible to address I2C memory (238)8.7.56 F2103 It was impossible to address EnDat memory (238)8.7.57 F2104 Commutation offset invalid (239)8.7.58 F2105 It was impossible to address Hiperface memory (239)8.7.59 F2110 Error in non-cyclical data communic. of power section (240)8.7.60 F2120 MMC: Defective or missing, replace (240)8.7.61 F2121 MMC: Incorrect data or file, create correctly (241)8.7.62 F2122 MMC: Incorrect IBF file, correct it (241)8.7.63 F2123 Retain data backup impossible (242)8.7.64 F2124 MMC: Saving too slowly, replace (243)8.7.65 F2130 Error comfort control panel (243)8.7.66 F2140 CCD slave error (243)8.7.67 F2150 MLD motion function block error (244)8.7.68 F2174 Loss of motor encoder reference (244)8.7.69 F2175 Loss of optional encoder reference (245)Troubleshooting Guide | Rexroth IndraDrive Electric Drivesand Controls| Bosch Rexroth AG IX/XXIITable of ContentsPage8.7.70 F2176 Loss of measuring encoder reference (246)8.7.71 F2177 Modulo limitation error of motor encoder (246)8.7.72 F2178 Modulo limitation error of optional encoder (247)8.7.73 F2179 Modulo limitation error of measuring encoder (247)8.7.74 F2190 Incorrect Ethernet configuration (248)8.7.75 F2260 Command current limit shutoff (249)8.7.76 F2270 Analog input 1 or 2, wire break (249)8.7.77 F2802 PLL is not synchronized (250)8.7.78 F2814 Undervoltage in mains (250)8.7.79 F2815 Overvoltage in mains (251)8.7.80 F2816 Softstart fault power supply unit (251)8.7.81 F2817 Overvoltage in power section (251)8.7.82 F2818 Phase failure (252)8.7.83 F2819 Mains failure (253)8.7.84 F2820 Braking resistor overload (253)8.7.85 F2821 Error in control of braking resistor (254)8.7.86 F2825 Switch-on threshold braking resistor too low (255)8.7.87 F2833 Ground fault in motor line (255)8.7.88 F2834 Contactor control error (256)8.7.89 F2835 Mains contactor wiring error (256)8.7.90 F2836 DC bus balancing monitor error (257)8.7.91 F2837 Contactor monitoring error (257)8.7.92 F2840 Error supply shutdown (257)8.7.93 F2860 Overcurrent in mains-side power section (258)8.7.94 F2890 Invalid device code (259)8.7.95 F2891 Incorrect interrupt timing (259)8.7.96 F2892 Hardware variant not supported (259)8.8 SERCOS Error Codes / Error Messages of Serial Communication (259)9 Warnings (Exxxx) (263)9.1 Fatal Warnings (E8xxx) (263)9.1.1 Behavior in the Case of Fatal Warnings (263)9.1.2 E8025 Overvoltage in power section (263)9.1.3 E8026 Undervoltage in power section (264)9.1.4 E8027 Safe torque off while drive enabled (265)9.1.5 E8028 Overcurrent in power section (265)9.1.6 E8029 Positive position limit exceeded (266)9.1.7 E8030 Negative position limit exceeded (267)9.1.8 E8034 Emergency-Stop (268)9.1.9 E8040 Torque/force actual value limit active (268)9.1.10 E8041 Current limit active (269)9.1.11 E8042 Both travel range limit switches activated (269)9.1.12 E8043 Positive travel range limit switch activated (270)9.1.13 E8044 Negative travel range limit switch activated (271)9.1.14 E8055 Motor overload, current limit active (271)9.1.15 E8057 Device overload, current limit active (272)X/XXII Bosch Rexroth AG | Electric Drivesand ControlsRexroth IndraDrive | Troubleshooting GuideTable of ContentsPage9.1.16 E8058 Drive system not ready for operation (273)9.1.17 E8260 Torque/force command value limit active (273)9.1.18 E8802 PLL is not synchronized (274)9.1.19 E8814 Undervoltage in mains (275)9.1.20 E8815 Overvoltage in mains (275)9.1.21 E8818 Phase failure (276)9.1.22 E8819 Mains failure (276)9.2 Warnings of Category E4xxx (277)9.2.1 E4001 Double MST failure shutdown (277)9.2.2 E4002 Double MDT failure shutdown (278)9.2.3 E4005 No command value input via master communication (279)9.2.4 E4007 SERCOS III: Consumer connection failed (280)9.2.5 E4008 Invalid addressing command value data container A (280)9.2.6 E4009 Invalid addressing actual value data container A (281)9.2.7 E4010 Slave not scanned or address 0 (281)9.2.8 E4012 Maximum number of CCD slaves exceeded (282)9.2.9 E4013 Incorrect CCD addressing (282)9.2.10 E4014 Incorrect phase switch of CCD slaves (283)9.3 Possible Warnings When Operating Safety Technology (E3xxx) (283)9.3.1 Behavior in Case a Safety Technology Warning Occurs (283)9.3.2 E3100 Error when checking input signals (284)9.3.3 E3101 Error when checking acknowledgment signal (284)9.3.4 E3102 Actual position values validation error (285)9.3.5 E3103 Dynamization failed (285)9.3.6 E3104 Safety parameters validation error (286)9.3.7 E3105 Validation error of safe operation mode (286)9.3.8 E3106 System error safety technology (287)9.3.9 E3107 Safe reference missing (287)9.3.10 E3108 Safely-monitored deceleration exceeded (288)9.3.11 E3110 Time interval of forced dynamization exceeded (289)9.3.12 E3115 Prewarning, end of brake check time interval (289)9.3.13 E3116 Nominal load torque of holding system reached (290)9.4 Non-Fatal Warnings (E2xxx) (290)9.4.1 Behavior in Case a Non-Fatal Warning Occurs (290)9.4.2 E2010 Position control with encoder 2 not possible (291)9.4.3 E2011 PLC - Warning no. 1 (291)9.4.4 E2012 PLC - Warning no. 2 (291)9.4.5 E2013 PLC - Warning no. 3 (292)9.4.6 E2014 PLC - Warning no. 4 (292)9.4.7 E2021 Motor temperature outside of measuring range (292)9.4.8 E2026 Undervoltage in power section (293)9.4.9 E2040 Device overtemperature 2 prewarning (294)9.4.10 E2047 Interpolation velocity = 0 (294)9.4.11 E2048 Interpolation acceleration = 0 (295)9.4.12 E2049 Positioning velocity >= limit value (296)9.4.13 E2050 Device overtemp. Prewarning (297)Troubleshooting Guide | Rexroth IndraDrive Electric Drivesand Controls| Bosch Rexroth AG XI/XXIITable of ContentsPage9.4.14 E2051 Motor overtemp. prewarning (298)9.4.15 E2053 Target position out of travel range (298)9.4.16 E2054 Not homed (300)9.4.17 E2055 Feedrate override S-0-0108 = 0 (300)9.4.18 E2056 Torque limit = 0 (301)9.4.19 E2058 Selected positioning block has not been programmed (302)9.4.20 E2059 Velocity command value limit active (302)9.4.21 E2061 Device overload prewarning (303)9.4.22 E2063 Velocity command value > limit value (304)9.4.23 E2064 Target position out of num. range (304)9.4.24 E2069 Holding brake torque too low (305)9.4.25 E2070 Acceleration limit active (306)9.4.26 E2074 Encoder 1: Encoder signals disturbed (306)9.4.27 E2075 Encoder 2: Encoder signals disturbed (307)9.4.28 E2076 Measuring encoder: Encoder signals disturbed (308)9.4.29 E2077 Absolute encoder monitoring, motor encoder (encoder alarm) (308)9.4.30 E2078 Absolute encoder monitoring, opt. encoder (encoder alarm) (309)9.4.31 E2079 Absolute enc. monitoring, measuring encoder (encoder alarm) (309)9.4.32 E2086 Prewarning supply module overload (310)9.4.33 E2092 Internal synchronization defective (310)9.4.34 E2100 Positioning velocity of master axis generator too high (311)9.4.35 E2101 Acceleration of master axis generator is zero (312)9.4.36 E2140 CCD error at node (312)9.4.37 E2270 Analog input 1 or 2, wire break (312)9.4.38 E2802 HW control of braking resistor (313)9.4.39 E2810 Drive system not ready for operation (314)9.4.40 E2814 Undervoltage in mains (314)9.4.41 E2816 Undervoltage in power section (314)9.4.42 E2818 Phase failure (315)9.4.43 E2819 Mains failure (315)9.4.44 E2820 Braking resistor overload prewarning (316)9.4.45 E2829 Not ready for power on (316)。

ADAPTIVE LASSO FOR SPARSE HIGH-DIMENSIONAL REGRESSION MODELS

in sparse, high-dimensional, linear regression models when the number of covariates may increase with the sample size. We consider variable selection using the adaptive Lasso, where the L1 norms in the penalty are re-weighted by data-dependent weights. We show that, if a reasonable initial estimator is available, then under appropriate conditions, the adaptive Lasso correctly selects covariates with nonzero coeﬃcients with probability converging to one and that the estimators of nonzero coeﬃcients have the same asymptotic distribution that they would have if the zero coeﬃcients were known in advance. Thus, the adaptive Lasso has an oracle property in the sense of Fan and Li (2001) and Fan and Peng (2004). In addition, under a partial orthogonality condition in which the covariates with zero coeﬃcients are weakly correlated with the covariates with nonzero coeﬃcients, marginal regression can be used to obtain the initial estimator. With this initial estimator, adaptive Lasso has the oracle property even when the number of covariates is much larger than the sample size. Key Words and phrases. Penalized regression, high-dimensional data, variable selection, asymptotic normality, oracle property, zero-consistency. Short title. Adaptive Lasso AMS 2000 subject classiﬁcation. Primary 62J05, 62J07; secondary 62E20, 60F05

Shimadzu - LCMS-2020 Vacuum System Main Unit ESI P

6 month Service X
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36 month Service 48 month Service 60 month Service
X X
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Note: These time frames are only suggestions and items may need to be replaced more frequently depending on sample throughput and use
X
Note: These time frames are only suggestions and items may need to be replaced more frequently depending on sample throughput and use
Shop online ~ or call Shimadzu at 800-477-1227
Shop online ~ or call Shimadzu at 800-477-1227
2
LCMS-8030/8040
Triple Quadrupole Mass Spectrometer
LCMS-8030
LCMS-8040
Vacuum System
Main Unit
SSI-LCMS-11-2015
Liquid Chromatography Mass Spectrometry
LCMS Consumables
LCMS-2020
Single Quadrupole Mass Spectrometer
Inspection/Maintenance Schedule

1999.Multilevel Hypergraph Partitioning__Applications in VLSI Domain

Multilevel Hypergraph Partitioning:Applications in VLSI DomainGeorge Karypis,Rajat Aggarwal,Vipin Kumar,Senior Member,IEEE,and Shashi Shekhar,Senior Member,IEEE Abstract—In this paper,we present a new hypergraph-partitioning algorithm that is based on the multilevel paradigm.In the multilevel paradigm,a sequence of successivelycoarser hypergraphs is constructed.A bisection of the smallesthypergraph is computed and it is used to obtain a bisection of theoriginal hypergraph by successively projecting and reﬁning thebisection to the next levelﬁner hypergraph.We have developednew hypergraph coarsening strategies within the multilevelframework.We evaluate their performance both in terms of thesize of the hyperedge cut on the bisection,as well as on the runtime for a number of very large scale integration circuits.Ourexperiments show that our multilevel hypergraph-partitioningalgorithm produces high-quality partitioning in a relatively smallamount of time.The quality of the partitionings produced by ourscheme are on the average6%–23%better than those producedby other state-of-the-art schemes.Furthermore,our partitioningalgorithm is signiﬁcantly faster,often requiring4–10times lesstime than that required by the other schemes.Our multilevelhypergraph-partitioning algorithm scales very well for largehypergraphs.Hypergraphs with over100000vertices can bebisected in a few minutes on today’s workstations.Also,on thelarge hypergraphs,our scheme outperforms other schemes(inhyperedge cut)quite consistently with larger margins(9%–30%).Index Terms—Circuit partitioning,hypergraph partitioning,multilevel algorithms.I.I NTRODUCTIONH YPERGRAPH partitioning is an important problem withextensive application to many areas,including very largescale integration(VLSI)design[1],efﬁcient storage of largedatabases on disks[2],and data mining[3].The problemis to partition the vertices of a hypergraphintois deﬁnedas a set ofvertices[4],and the size ofa hyperedge is the cardinality of this subset.Manuscript received April29,1997;revised March23,1998.This workwas supported under IBM Partnership Award NSF CCR-9423082,by theArmy Research Ofﬁce under Contract DA/DAAH04-95-1-0538,and by theArmy High Performance Computing Research Center,the Department of theArmy,Army Research Laboratory Cooperative Agreement DAAH04-95-2-0003/Contract DAAH04-95-C-0008.G.Karypis,V.Kumar,and S.Shekhar are with the Department of ComputerScience and Engineering,Minneapolis,University of Minnesota,Minneapolis,MN55455-0159USA.R.Aggarwal is with the Lattice Semiconductor Corporation,Milpitas,CA95131USA.Publisher Item Identiﬁer S1063-8210(99)00695-2.During the course of VLSI circuit design and synthesis,itis important to be able to divide the system speciﬁcation intoclusters so that the inter-cluster connections are minimized.This step has many applications including design packaging,HDL-based synthesis,design optimization,rapid prototyping,simulation,and testing.In particular,many rapid prototyp-ing systems use partitioning to map a complex circuit ontohundreds of interconnectedﬁeld-programmable gate arrays(FPGA’s).Such partitioning instances are challenging becausethe timing,area,and input/output(I/O)resource utilizationmust satisfy hard device-speciﬁc constraints.For example,ifthe number of signal nets leaving any one of the clustersis greater than the number of signal p-i-n’s available in theFPGA,then this cluster cannot be implemented using a singleFPGA.In this case,the circuit needs to be further partitioned,and thus implemented using multiple FPGA’s.Hypergraphscan be used to naturally represent a VLSI circuit.The verticesof the hypergraph can be used to represent the cells of thecircuit,and the hyperedges can be used to represent the netsconnecting these cells.A high quality hypergraph-partitioningalgorithm greatly affects the feasibility,quality,and cost ofthe resulting system.A.Related WorkThe problem of computing an optimal bisection of a hy-pergraph is at least NP-hard[5].However,because of theimportance of the problem in many application areas,manyheuristic algorithms have been developed.The survey byAlpert and Khang[1]provides a detailed description andcomparison of such various schemes.In a widely used class ofiterative reﬁnement partitioning algorithms,an initial bisectionis computed(often obtained randomly)and then the partitionis reﬁned by repeatedly moving vertices between the twoparts to reduce the hyperedge cut.These algorithms oftenuse the Schweikert–Kernighan heuristic[6](an extension ofthe Kernighan–Lin(KL)heuristic[7]for hypergraphs),or thefaster Fiduccia–Mattheyses(FM)[8]reﬁnement heuristic,toiteratively improve the quality of the partition.In all of thesemethods(sometimes also called KLFM schemes),a vertex ismoved(or a vertex pair is swapped)if it produces the greatestreduction in the edge cuts,which is also called the gain formoving the vertex.The partition produced by these methodsis often poor,especially for larger hypergraphs.Hence,thesealgorithms have been extended in a number of ways[9]–[12].Krishnamurthy[9]tried to introduce intelligence in the tie-breaking process from among the many possible moves withthe same high gain.He used a Look Ahead()algorithm,which looks ahead uptoa move.PROP [11],introduced by Dutt and Deng,used a probabilistic gain computation model for deciding which vertices need to move across the partition line.These schemes tend to enhance the performance of the basic KLFM family of reﬁnement algorithms,at the expense of increased run time.Dutt and Deng [12]proposed two new methods,namely,CLIP and CDIP,for computing the gains of hyperedges that contain more than one node on either side of the partition boundary.CDIP in conjunctionwithand CLIP in conjunction with PROP are two schemes that have shown the best results in their experiments.Another class of hypergraph-partitioning algorithms [13]–[16]performs partitioning in two phases.In the ﬁrst phase,the hypergraph is coarsened to form a small hypergraph,and then the FM algorithm is used to bisect the small hypergraph.In the second phase,these algorithms use the bisection of this contracted hypergraph to obtain a bisection of the original hypergraph.Since FM reﬁnement is done only on the small coarse hypergraph,this step is usually fast,but the overall performance of such a scheme depends upon the quality of the coarsening method.In many schemes,the projected partition is further improved using the FM reﬁnement scheme [15].Recently,a new class of partitioning algorithms was devel-oped [17]–[20]based upon the multilevel paradigm.In these algorithms,a sequence of successively smaller (coarser)graphs is constructed.A bisection of the smallest graph is computed.This bisection is now successively projected to the next-level ﬁner graph and,at each level,an iterative reﬁnement algorithm such as KLFM is used to further improve the bisection.The various phases of multilevel bisection are illustrated in Fig.1.Iterative reﬁnement schemes such as KLFM become quite powerful in this multilevel context for the following reason.First,the movement of a single node across a partition bound-ary in a coarse graph can lead to the movement of a large num-ber of related nodes in the original graph.Second,the reﬁned partitioning projected to the next level serves as an excellent initial partitioning for the KL or FM reﬁnement algorithms.This paradigm was independently studied by Bui and Jones [17]in the context of computing ﬁll-reducing matrix reorder-ing,by Hendrickson and Leland [18]in the context of ﬁnite-element mesh-partitioning,and by Hauck and Borriello (called Optimized KLFM)[20],and by Cong and Smith [19]for hy-pergraph partitioning.Karypis and Kumar extensively studied this paradigm in [21]and [22]for the partitioning of graphs.They presented new graph coarsening schemes for which even a good bisection of the coarsest graph is a pretty good bisec-tion of the original graph.This makes the overall multilevel paradigm even more robust.Furthermore,it allows the use of simpliﬁed variants of KLFM reﬁnement schemes during the uncoarsening phase,which signiﬁcantly speeds up the reﬁne-ment process without compromising overall quality.METIS [21],a multilevel graph partitioning algorithm based upon this work,routinely ﬁnds substantially better bisections and is often two orders of magnitude faster than the hitherto state-of-the-art spectral-based bisection techniques [23],[24]for graphs.The improved coarsening schemes of METIS work only for graphs and are not directly applicable to hypergraphs.IftheFig.1.The various phases of the multilevel graph bisection.During the coarsening phase,the size of the graph is successively decreased;during the initial partitioning phase,a bisection of the smaller graph is computed,and during the uncoarsening and reﬁnement phase,the bisection is successively reﬁned as it is projected to the larger graphs.During the uncoarsening and reﬁnement phase,the dashed lines indicate projected partitionings and dark solid lines indicate partitionings that were produced after reﬁnement.G 0is the given graph,which is the ﬁnest graph.G i +1is the next level coarser graph of G i ,and vice versa,G i is the next level ﬁner graph of G i +1.G 4is the coarsest graph.hypergraph is ﬁrst converted into a graph (by replacing each hyperedge by a set of regular edges),then METIS [21]can be used to compute a partitioning of this graph.This technique was investigated by Alpert and Khang [25]in their algorithm called GMetis.They converted hypergraphs to graphs by simply replacing each hyperedge with a clique,and then they dropped many edges from each clique randomly.They used METIS to compute a partitioning of each such random graph and then selected the best of these partitionings.Their results show that reasonably good partitionings can be obtained in a reasonable amount of time for a variety of benchmark problems.In particular,the performance of their resulting scheme is comparable to other state-of-the art schemes such as PARABOLI [26],PROP [11],and the multilevel hypergraph partitioner from Hauck and Borriello [20].The conversion of a hypergraph into a graph by replacing each hyperedge with a clique does not result in an equivalent representation since high-quality partitionings of the resulting graph do not necessarily lead to high-quality partitionings of the hypergraph.The standard hyperedge-to-edge conversion [27]assigns a uniform weightofisthe of the hyperedge,i.e.,thenumber of vertices in the hyperedge.However,the fundamen-tal problem associated with replacing a hyperedge by its clique is that there exists no scheme to assign weight to the edges of the clique that can correctly capture the cost of cutting this hyperedge [28].This hinders the partitioning reﬁnement algo-rithm since vertices are moved between partitions depending on how much this reduces the number of edges they cut in the converted graph,whereas the real objective is to minimize the number of hyperedges cut in the original hypergraph.Furthermore,the hyperedge-to-clique conversion destroys the natural sparsity of the hypergraph,signiﬁcantly increasing theKARYPIS et al.:MULTILEVEL HYPERGRAPH PARTITIONING:APPLICATIONS IN VLSI DOMAIN 71run time of the partitioning algorithm.Alpert and Khang [25]solved this problem by dropping many edges of the clique randomly,but this makes the graph representation even less accurate.A better approach is to develop coarsening and reﬁnement schemes that operate directly on the hypergraph.Note that the multilevel scheme by Hauck and Borriello [20]operates directly on hypergraphs and,thus,is able to perform accurate reﬁnement during the uncoarsening phase.However,all coarsening schemes studied in [20]are edge-oriented;i.e.,they only merge pairs of nodes to construct coarser graphs.Hence,despite a powerful reﬁnement scheme (FM with theuse oflook-ahead)during the uncoarsening phase,their performance is only as good as that of GMetis [25].B.Our ContributionsIn this paper,we present a multilevel hypergraph-partitioning algorithm hMETIS that operates directly on the hypergraphs.A key contribution of our work is the development of new hypergraph coarsening schemes that allow the multilevel paradigm to provide high-quality partitions quite consistently.The use of these powerful coarsening schemes also allows the reﬁnement process to be simpliﬁed considerably (even beyond plain FM reﬁnement),making the multilevel scheme quite fast.We investigate various algorithms for the coarsening and uncoarsening phases which operate on the hypergraphs without converting them into graphs.We have also developed new multiphase reﬁnement schemes(-cycles)based on the multilevel paradigm.These schemes take an initial partition as input and try to improve them using the multilevel scheme.These multiphase schemes further reduce the run times,as well as improve the solution quality.We evaluate their performance both in terms of the size of the hyperedge cut on the bisection,as well as on run time on a number of VLSI circuits.Our experiments show that our multilevel hypergraph-partitioning algorithm produces high-quality partitioning in a relatively small amount of time.The quality of the partitionings produced by our scheme are on the average 6%–23%better than those produced by other state-of-the-art schemes [11],[12],[25],[26],[29].The difference in quality over other schemes becomes even greater for larger hypergraphs.Furthermore,our partitioning algorithm is signiﬁcantly faster,often requiring 4–10times less time than that required by the other schemes.For many circuits in the well-known ACM/SIGDA benchmark set [30],our scheme is able to ﬁnd better partitionings than those reported in the literature for any other hypergraph-partitioning algorithm.The remainder of this paper is organized as follows.Section II describes the different algorithms used in the three phases of our multilevel hypergraph-partitioning algorithm.Section III describes a new partitioning reﬁnement algorithm based on the multilevel paradigm.Section IV compares the results produced by our algorithm to those produced by earlier hypergraph-partitioning algorithms.II.M ULTILEVEL H YPERGRAPH B ISECTIONWe now present the framework of hMETIS ,in which the coarsening and reﬁnement scheme work directly with hyper-edges without using the clique representation to transform them into edges.We have developed new algorithms for both the phases,which,in conjunction,are capable of delivering very good quality solutions.A.Coarsening PhaseDuring the coarsening phase,a sequence of successively smaller hypergraphs are constructed.As in the case of mul-tilevel graph bisection,the purpose of coarsening is to create a small hypergraph,such that a good bisection of the small hypergraph is not signiﬁcantly worse than the bisection di-rectly obtained for the original hypergraph.In addition to that,hypergraph coarsening also helps in successively reducing the sizes of the hyperedges.That is,after several levels of coarsening,large hyperedges are contracted to hyperedges that connect just a few vertices.This is particularly helpful,since reﬁnement heuristics based on the KLFM family of algorithms [6]–[8]are very effective in reﬁning small hyperedges,but are quite ineffective in reﬁning hyperedges with a large number of vertices belonging to different partitions.Groups of vertices that are merged together to form single vertices in the next-level coarse hypergraph can be selected in different ways.One possibility is to select pairs of vertices with common hyperedges and to merge them together,as illustrated in Fig.2(a).A second possibility is to merge together all the vertices that belong to a hyperedge,as illustrated in Fig.2(b).Finally,a third possibility is to merge together a subset of the vertices belonging to a hyperedge,as illustrated in Fig.2(c).These three different schemes for grouping vertices together for contraction are described below.1)Edge Coarsening (EC):The heavy-edge matching scheme used in the multilevel-graph bisection algorithm can also be used to obtain successively coarser hypergraphs by merging the pairs of vertices connected by many hyperedges.In this EC scheme,a heavy-edge maximal 1matching of the vertices of the hypergraph is computed as follows.The vertices are visited in a random order.For eachvertex are considered,and the one that is connected via the edge with the largest weight is matchedwithandandofsize72IEEE TRANSACTIONS ON VERY LARGE SCALE INTEGRATION(VLSI)SYSTEMS,VOL.7,NO.1,MARCH1999Fig.2.Various ways of matching the vertices in the hypergraph and the coarsening they induce.(a)In edge-coarsening,connected pairs of vertices are matched together.(b)In hyperedge-coarsening,all the vertices belonging to a hyperedge are matched together.(c)In MHEC,we match together all the vertices in a hyperedge,as well as all the groups of vertices belonging to a hyperedge.weight of successively coarser graphs does not decrease very fast.In order to ensure that for every group of vertices that are contracted together,there is a decrease in the hyperedge weight in the coarser graph,each such group of vertices must be connected by a hyperedge.This is the motivation behind the HEC scheme.In this scheme,an independent set of hyperedges is selected and the vertices that belong to individual hyperedges are contracted together.This is implemented as follows.The hyperedges are initially sorted in a nonincreasing hyperedge-weight order and the hyperedges of the same weight are sorted in a nondecreasing hyperedge size order.Then,the hyperedges are visited in that order,and for each hyperedge that connects vertices that have not yet been matched,the vertices are matched together.Thus,this scheme gives preference to the hyperedges that have large weight and those that are of small size.After all of the hyperedges have been visited,the groups of vertices that have been matched are contracted together to form the next level coarser graph.The vertices that are not part of any contracted hyperedges are simply copied to the next level coarser graph.3)Modiﬁed Hyperedge Coarsening(MHEC):The HEC algorithm is able to signiﬁcantly reduce the amount of hyperedge weight that is left exposed in successively coarser graphs.However,during each coarsening phase,a majority of the hyperedges do not get contracted because vertices that belong to them have been contracted via other hyperedges. This leads to two problems.First,the size of many hyperedges does not decrease sufﬁciently,making FM-based reﬁnement difﬁcult.Second,the weight of the vertices(i.e.,the number of vertices that have been collapsed together)in successively coarser graphs becomes signiﬁcantly different,which distorts the shape of the contracted hypergraph.To correct this problem,we implemented a MHEC scheme as follows.After the hyperedges to be contracted have been selected using the HEC scheme,the list of hyperedges is traversed again,and for each hyperedge that has not yet been contracted,the vertices that do not belong to any other contracted hyperedge are contracted together.B.Initial Partitioning PhaseDuring the initial partitioning phase,a bisection of the coarsest hypergraph is computed,such that it has a small cut, and satisﬁes a user-speciﬁed balance constraint.The balance constraint puts an upper bound on the difference between the relative size of the two partitions.Since this hypergraph has a very small number of vertices(usually less than200),the time toﬁnd a partitioning using any of the heuristic algorithms tends to be small.Note that it is not useful toﬁnd an optimal partition of this coarsest graph,as the initial partition will be sub-stantially modiﬁed during the reﬁnement phase.We used the following two algorithms for computing the initial partitioning. Theﬁrst algorithm simply creates a random bisection such that each part has roughly equal vertex weight.The second algorithm starts from a randomly selected vertex and grows a region around it in a breadth-ﬁrst fashion[22]until half of the vertices are in this region.The vertices belonging to the grown region are then assigned to theﬁrst part,and the rest of the vertices are assigned to the second part.After a partitioning is constructed using either of these algorithms,the partitioning is reﬁned using the FM reﬁnement algorithm.Since both algorithms are randomized,different runs give solutions of different quality.For this reason,we perform a small number of initial partitionings.At this point,we can select the best initial partitioning and project it to the original hypergraph,as described in Section II-C.However,the parti-tioning of the coarsest hypergraph that has the smallest cut may not necessarily be the one that will lead to the smallest cut in the original hypergraph.It is possible that another partitioning of the coarsest hypergraph(with a higher cut)will lead to a bet-KARYPIS et al.:MULTILEVEL HYPERGRAPH PARTITIONING:APPLICATIONS IN VLSI DOMAIN 73ter partitioning of the original hypergraph after the reﬁnement is performed during the uncoarsening phase.For this reason,instead of selecting a single initial partitioning (i.e.,the one with the smallest cut),we propagate all initial partitionings.Note that propagation of.Thus,by increasing the value ofis to drop unpromising partitionings as thehypergraph is uncoarsened.For example,one possibility is to propagate only those partitionings whose cuts arewithinissufﬁciently large,then all partitionings will be maintained and propagated in the entire reﬁnement phase.On the other hand,if the valueof,many partitionings may be available at the coarsest graph,but the number of such available partitionings will decrease as the graph is uncoarsened.This is useful for two reasons.First,it is more important to have many alternate partitionings at the coarser levels,as the size of the cut of a partitioning at a coarse level is a less accurate reﬂection of the size of the cut of the original ﬁnest level hypergraph.Second,reﬁnement is more expensive at the ﬁne levels,as these levels contain far more nodes than the coarse levels.Hence,by choosing an appropriate valueof(from 10%to a higher value such as 20%)did not signiﬁcantly improve the quality of the partitionings,although it did increase the run time.C.Uncoarsening and Reﬁnement PhaseDuring the uncoarsening phase,a partitioning of the coarser hypergraph is successively projected to the next-level ﬁner hypergraph,and a partitioning reﬁnement algorithm is used to reduce the cut set (and thus to improve the quality of the partitioning)without violating the user speciﬁed balance con-straints.Since the next-level ﬁner hypergraph has more degrees of freedom,such reﬁnement algorithms tend to improve the solution quality.We have implemented two different partitioning reﬁnement algorithms.The ﬁrst is the FM algorithm [8],which repeatedly moves vertices between partitions in order to improve the cut.The second algorithm,called hyperedge reﬁnement (HER),moves groups of vertices between partitions so that an entire hyperedge is removed from the cut.These algorithms are further described in the remainder of this section.1)FM:The partitioning reﬁnement algorithm by Fiduccia and Mattheyses [8]is iterative in nature.It starts with an initial partitioning of the hypergraph.In each iteration,it tries to ﬁnd subsets of vertices in each partition,such that moving them to other partitions improves the quality of the partitioning (i.e.,the number of hyperedges being cut decreases)and this does not violate the balance constraint.If such subsets exist,then the movement is performed and this becomes the partitioning for the next iteration.The algorithm continues by repeating the entire process.If it cannot ﬁnd such a subset,then the algorithm terminates since the partitioning is at a local minima and no further improvement can be made by this algorithm.In particular,for eachvertexto the other partition.Initially allvertices are unlocked ,i.e.,they are free to move to the other partition.The algorithm iteratively selects an unlockedvertex is moved,it is locked ,and the gain of the vertices adjacentto[8].For reﬁnement in the context of multilevel schemes,the initial partitioning obtained from the next level coarser graph is actually a very good partition.For this reason,we can make a number of optimizations to the original FM algorithm.The ﬁrst optimization limits the maximum number of passes performed by the FM algorithm to only two.This is because the greatest reduction in the cut is obtained during the ﬁrst or second pass and any subsequent passes only marginally improve the quality.Our experience has shown that this optimization signiﬁcantly improves the run time of FM without affecting the overall quality of the produced partitionings.The second optimization aborts each pass of the FM algorithm before actually moving all the vertices.The motivation behind this is that only a small fraction of the vertices being moved actually lead to a reduction in the cut and,after some point,the cut tends to increase as we move more vertices.When FM is applied to a random initial partitioning,it is quite likely that after a long sequence of bad moves,the algorithm will climb74IEEE TRANSACTIONS ON VERY LARGE SCALE INTEGRATION (VLSI)SYSTEMS,VOL.7,NO.1,MARCH1999Fig.3.Effect of restricted coarsening .(a)Example hypergraph with a given partitioning with the required balance of 40/60.(b)Possible condensed version of (a).(c)Another condensed version of a hypergraph.out of a local minima and reach to a better cut.However,in the context of a multilevel scheme,a long sequence of cut-increasing moves rarely leads to a better local minima.For this reason,we stop each pass of the FM algorithm as soon as we haveperformedto be equal to 1%of the number ofvertices in the graph we are reﬁning.This modiﬁcation to FM,called early-exit FM (FM-EE),does not signiﬁcantly affect the quality of the ﬁnal partitioning,but it dramatically improves the run time (see Section IV).2)HER:One of the drawbacks of FM (and other similar vertex-based reﬁnement schemes)is that it is often unable to reﬁne hyperedges that have many nodes on both sides of the partitioning boundary.However,a reﬁnement scheme that moves all the vertices that belong to a hyperedge can potentially solve this problem.Our HER works as follows.It randomly visits all the hyperedges and,for each one that straddles the bisection,it determines if it can move a subset of the vertices incident on it,so that this hyperedge will become completely interior to a partition.In particular,consider ahyperedgebe the verticesofto partition 0.Now,depending on these gains and subject to balance constraints,it may move one of the twosets .In particular,if.III.M ULTIPHASE R EFINEMENT WITHR ESTRICTED C OARSENINGAlthough the multilevel paradigm is quite robust,random-ization is inherent in all three phases of the algorithm.In particular,the random choice of vertices to be matched in the coarsening phase can disallow certain hyperedge cuts,reducing reﬁnement in the uncoarsening phase.For example,consider the example hypergraph in Fig.3(a)and its two possible con-densed versions [Fig.3(b)and (c)]with the same partitioning.The version in Fig.3(b)is obtained by selectinghyperedgesto be compressed in the HEC phase and then selecting pairs ofnodesto be compressed inthe HEC phase and then selecting pairs ofnodesand apartitioningfor ,be the sequence of hypergraphsand partitionings.Given ahypergraphandorpartition,,are collapsedtogether to formvertexof,thenvertex belong。

e Xpressway Integrator OnRamp 预设置检查列表说明书

e*Xpressway Integrator OnRampPre-setup ChecklistPrint this Checklist and fill it out. The information you provide will help you determineyour level of readiness to install and configure e*Xpressway Integrator OnRamp so youcan easily integrate with your Trading Exchange.Installation environment requirementsCheck One dedicated, clean PC (a clean PC has only the Operating System installed)IP address ____________________________________________________Note: A clean PC is strongly recommended but is not an absolute requirement. If youdecide not to start with a clean PC, you should first evaluate any potential performanceand/or capacity issues.Intel Pentium III or equivalent AMD Processor, 800Mhz minimumWindows 2000 with Service Pack 2 or NT 4.0 with Service Pack 6a or aboveMicrosoft Internet Explorer 5.0 or later with 128-bit encryption enabled512 Mb memory minimum1Gb of free disk space for installation and operational dataPersistent broadband Internet connectionConnection Type (T1, DSL, etc) _____________________________________Name of Internet Service Provider (ISP) _______________________________ISP 24-hour Hot Line number _______________________________________SecurityCertificate Authority URL _______________________________________Certificate location (required) ____________________________________Public Key location (required) ____________________________________Recommended Trusted Certificate AuthorityVeriSign is the recommended Trusted Certificate Authority for e*Xpressway Integrator OnRamp users with average computing experience. On their Web site, VeriSign offersstep-by-step instructions for acquiring a Certificate and Private Key.e*Xpressway Integrator OnRamp requires 128-bit SSL encryption. If your current SSLencryption level is not at 128-bit, you must purchase an upgrade.Note: The steps below are correct as of the date of this release of e*Xpressway Integrator OnRamp. However, these steps can be invalidated by interim changes made to theVeriSign Web site.To acquire the recommended Certificate and Key, do the following1. From your browser, go to .2. On the main menu banner at the top of the page, select Products.3. Under the Retail Services heading, click the SSL Certificates link.4. Under the Secure Site Services heading, click Buy.5. Follow the purchasing instructions on the Web site.Additional Trusted Certificate AuthoritiesThe following Trusted Certificates are also supported by e*Xpressway IntegratorOnRamp, but step-by-step instructions are NOT offered on their Web sites, making itmuch more difficult for users with limited-to-average computing experience to acquire a Certificate and Key:GETCyberTrustGlobalRoot.cer ( )MicrosoftRootAuthority.cer ()SecureServerCertificationAuthority.cer ()ThawtePremiumServerCA.cer ()ThawteServerCA.cer ()Human resources: Integration ManagerName __________________________________________________Contact Number __________________________________________ExperienceFull access System Administrator privileges (Yes/No) ________________________ Knowledge of network email systems (Yes/No)Experience with MS Internet Explorer (Yes/No) ____________Knowledge of Internet security technology (Yes/No) ____________Note: It is strongly recommended that you designate and train a backup IntegrationManager.Information requirementsDomain Name for your Web ServerDomain ________________________________________________URL for your Trading Partner serverNote: This URL will be provided when you install e*Xpressway Integrator OnRamp. Fill this line in when the URL is displayed during the installation procedure.URL ___________________________________________________ Email address used for logon to membership area on Trading ExchangeAdministrator ____________________________________________Back office file format(s) for B2B messages (for OnRamp Solution Packages)Format ________________________________________Format ________________________________________Format ________________________________________Format ________________________________________For more information, refer to the e*Xpressway Integrator OnRamp Setup Guide forTrading Partners available in PDF format on the Trading Exchange Web site.© 2001 by SeeBeyond Technology Corporation. All Rights Reserved. This work is protected as an unpublished work under the copyright laws.。

版纳微型猪近交系TNF-α基因mRNA实时荧光定量PCR检测方法的建立

ｆｄｐｏｕｔｏＰＲｗｒｏｊｇｔｄｗｔａｐ１－ｅｔｎｒｎｆｒｄｉｏｔｅｂｃｒＨａｆｅｌａｉ．ＴｅｉｒｄｃｆＣｅｅｃｎｕａｉＭＤ８ＴｖｃｒｄｔｓｒｔｈａｔｉＤ５ｒｒｉｔｎｈｅｓｅｈｏａａｅｅｎｅａｏｐｃｏ
取皮肤创面总ＲＡ，计特异引物，行Ｒ．Ｃ扩增。纯化目的片段与ｐ１．体连接，化宿主菌Ｄ５Ｎ设进ＴＰＲＭＤ８Ｔ载转Ｈａ，提取重组质粒ＤＡ，经酶切、Ｃ和测序鉴定，算重组质粒原液拷贝数浓度并制备梯度浓度标准品，行实时Ｎ并ＰＲ计进荧光定量ＰＲ，成标准曲线。结果Ｃ生建立的孙一基因和ＧＰＨ内参基因ｍＲＡ表达实时荧光定量ＰＲ检测ＡＤＮＣ方法灵敏度分别可达ｌ。１拷贝，性范围分别为１～１’和１～１’ 贝，Ｏ和Ｏ线００００拷阈值循环数（ｔ与ＰＲ体系中Ｃ）Ｃ起始模板量的对数值之间存在的线性关系Ｒ分别为０９３和０９９扩增效率Ｅ分别为１１０３和９．４％。．９．９，１．７％５９８结论成功的构建了版纳微型猪近交系＂一基因质粒标准品和标准曲线，用内参基因ＧＰＨ进行校正，方Ｏｔ并ＡＤ此法可为探讨Ｆ基因在临床多种疾病中所发挥的分子机理奠定基础。 —

A Versatile Zero Background T-Vector System for Gene

Breakthrough TechnologiesA Versatile Zero Background T-Vector System for Gene Cloning and Functional Genomics1[C][W][OA]Songbiao Chen,Pattavipha Songkumarn,Jianli Liu,and Guo-Liang Wang*Department of Plant Pathology,The Ohio State University,Columbus,Ohio43210(S.C.,P.S.,J.L.,G.-L.W.); and Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization,Hunan Agricultural University,Changsha410128,China(G.-L.W.)With the recent availability of complete genomic sequences of many organisms,high-throughput and cost-efﬁcient systems for gene cloning and functional analysis are in great demand.Although site-speciﬁc recombination-based cloning systems,such as Gateway cloning technology,are extremely useful for efﬁcient transfer of DNA fragments into multiple destination vectors,the two-step cloning process is time consuming and expensive.Here,we report a zero background TA cloning system that provides simple and high-efﬁciency direct cloning of PCR-ampliﬁed DNA fragments with almost no self-ligation.The improved T-vector system takes advantage of the restriction enzyme Xcm I to generate a T-overhang after digestion and the negative selection marker gene ccdB to eliminate the self-ligation background after transformation.We demonstrate the feasibility andﬂexibility of the technology by developing a set of transient and stable transformation vectors for constitutive gene expression,gene silencing,protein tagging,protein subcellular localization detection,and promoter fragment activity analysis in plants.Because the system can be easily adapted for developing specialized expression vectors for other organisms, zero background TA provides a general,cost-efﬁcient,and high-throughput platform that complements the Gateway cloning system for gene cloning and functional genomics.Rapid advances in genome sequencing technologies in the last few years have led to the complete decoding of many complex eukaryotic genomes and have stim-ulated large-scale analysis of gene functions in se-quenced genomes.In general,gene function can be elucidated using a variety of approaches,such as ectopic expression,gene silencing,protein subcellular localization examination,gene expression pattern analysis by promoter activity assay,structure-function analysis,and in vitro or in vivo biochemical assays (Hartley et al.,2000;Curtis and Grossniklaus,2003; Earley et al.,2006).Typically,all these approaches require the cloning of target genes,mutated fragments, or their promoter fragments into various specialized vectors for subsequent characterization.However,the traditional approach for engineering expression con-structs based on the restriction enzyme/ligase cloning method is extremely laborious and time consuming and is often hampered by lack of appropriate restric-tion sites;thus,the production of constructs is a signiﬁcant technical obstacle for large-scale functional gene analysis in plants.In recent years,the Gateway cloning system from Invitrogen and the Creator cloning system from CLONTECH have been developed to facilitate large-scale production of gene constructs.The recombina-tional cloning systems are based on a two-step process (Marsischky and LaBaer,2004).The DNA fragment of interest isﬁrst cloned into a general donor plasmid. Subsequently,the DNA fragmentﬂanked by two site-speciﬁc recombination sites in the donor vector can be transferred precisely into a variety of expression vec-tors by site-speciﬁc recombination reactions.A great advantage of the recombinational cloning technologies is that once the DNA fragment has been engineered into a donor vector,the transfer of the DNA fragment into an expression destination vector is a simple reac-tion that requires no traditional restriction enzyme/ ligase cloning.The recombinational cloning systems, particularly the Gateway technology,have been widely used in the research community,and many Gateway-compatible open reading frame entry(do-nor)clone collections and expression vectors have been created for functional genomics in many organ-isms(Yashiroda et al.,2008),including plants(Karimi et al.,2007b).On the other hand,although extremely useful for the simple and efﬁcient transfer of DNA fragments into multiple expression destination vec-tors,the usefulness of the Gateway cloning system is rather limited for many projects where only a single expression vector is required for a DNA fragment.The two-step cloning process of the Gateway technology is laborious and time consuming for the production of a1This work was supported by the National Science Foundation-Plant Genome Research Program(grant no.0605017).*Corresponding author;e-mail wang.620@.The author responsible for distribution of materials integral to the ﬁndings presented in this article in accordance with the policy described in the Instructions for Authors()is: Guo-Liang Wang(wang.620@).[C]Someﬁgures in this article are displayed in color online but in black and white in the print edition.[W]The online version of this article contains Web-only data.[OA]Open access articles can be viewed online without a sub-scription./cgi/doi/10.1104/pp.109.137125single expression vector.This is particularly true when a large number of plasmids must be cloned.Although a one-step recombinational cloning method was de-scribed to eliminate the production of an entry clone (Fu et al.,2008),the approach is rather limited in scope because long primers containing the speciﬁc attach-ment site(att)and two-step PCR are required(Fu et al., 2008).TA cloning is routinely used for cloning of PCR-ampliﬁed fragments.This technique exploits the ter-minal transferase activity of some DNA polymerases that add a3#-A overhang to each end of the PCR product.PCR products can be easily cloned into a linearized vector with3#-T overhangs compatible with 3#-A overhangs.Because it is difﬁcult to generate a high-quality TA cloning vector in individual laborato-ries,many TA cloning kits are available in the market. Many of them use blue/white screening for recombi-nants,and the DNA fragments can only be cloned into the TA vector provided in the kit.To meet the need for high-throughput cloning of DNA fragments into di-verse expression vectors,we have developed a signif-icantly improved TA cloning vector system by taking advantage of the negative selection gene marker ccdB to eliminate the self-ligation background after trans-formation.We refer to this new method as the zero background TA cloning system(ZeBaTA).Numerous cloning tests in our laboratory have shown that ZeBaTA provides very high cloning efﬁciency with almost no self-ligation.Moreover,the ZeBaTA technology can be ﬂexibly adapted for developing specialized expression vectors allowing single-step assembly of PCR-ampli-ﬁed genes or fragments.We demonstrate the feasibility andﬂexibility of the technology by developing a set of 12transient and12stable transformation vectors for constitutive gene expression,gene silencing,protein tagging,protein subcellular localization,and promoter fragment activity analysis for rice(Oryza sativa)and Arabidopsis(Arabidopsis thaliana).Our results suggest that ZeBaTA technology can also be easily used to develop expression vectors for other organisms(e.g. Escherichia coli,yeast[Saccharomyces cerevisiae],insect, and mammal),thereby providing a novel and general high-throughput platform for functional genomics of target genes.RESULTSConstruction of the ZeBaTA SystemTwo different strategies were used to produce T-vectors,i.e.adding a single thymidine at the3# blunt ends of a linearized vector(Holton and Graham, 1991;Marchuk et al.,1991)and generating single3#-T overhangs of a linearized vector by restriction endo-nuclease digestion(Kovalic et al.,1991;Mead et al., 1991;Ichihara and Kurosawa,1993;Chen et al.,2006a). Although the former has been used to produce com-mercial cloning kits like the pGEM-T system,we selected the restriction endonuclease digestion-mediated strategy to develop a TA cloning vector system because this approach is easy to use for indi-vidual laboratories.Previous publications have de-scribed the use of restriction enzyme Xcm I(Kovalic et al.,1991;Mead et al.,1991)or Ahd I/Eam1105I (Ichihara and Kurosawa,1993;Chen et al.,2006a)to produce intermediate T-vectors.We chose Xcm I as the digestion enzyme to develop the ZeBaTA cloning system because it had a better digestion efﬁciency than AhdI.Figure1.Construction of the ZeBaTA system.A,Schematic represen-tation of direct cloning of PCR product using the ZeBaTA vector system. The linker of the vector(in gray)is removed after Xcm I digestion yielding a linearized T-vector.B,TA cloning tests of the ZeBaTA system.(1)Self-ligation of Xcm I-digested pGXT using T4DNA ligase from Promega.(2)Ligation of Xcm I-digested pGXT with the PCR product of the rice blast fungus M.oryzae gene MGG_07986.5using T4DNA ligase from Promega.(3)Ligation of Xcm I-digested pGXT with the PCR product of MGG_07986.5using T4DNA ligase from USB Corporation. C,Samples of restriction digestion analysis of the randomly selected colonies derived from ligation of Xcm I-digested pGXT with the PCR product of MGG_07986.5using T4DNA ligase from Promega.pGXT contains two Bam HI recognition sites outside the two Xcm I recognition sites(Supplemental Fig.S1),and MGG_07986.5contains one internal Bam HI site.All samples(lanes1–20)digested by Bam HI released two bands as expected.M,1-kb DNA ladder.Chen et al.The schematic illustration of the improved T-vector system for PCR-ampliﬁed gene/fragment cloning is shown in Figure 1A.A pair of Xcm I recognition sites,CCAATACT/TGTATGG,was introduced in the vec-tors,which allowed the generation of a single thymi-dine residue at both 3#ends of the vector when digested with Xcm I.To eliminate the potential self-ligation due to incomplete Xcm I digestion of the vec-tor,the ccdB gene (Bernard and Couturier,1992;Miki et al.,1992),which inhibits growth of E .coli strains by expressing a protein to interfere with its DNA gyrase,was introduced between the two Xcm I sites.Hence,any self-ligation transformants containing the ccdB gene will be eliminated.To test the cloning efﬁciency of the T-vector system,an intermediate vector pGXT was generated based on the backbone of the pGEM-T easy vector.After Xcm I digestion,ligation reactions of the resulting T-vector alone and T-vector with the PCR-ampliﬁed product of the rice blast fungus Mag-naporthe oryzae gene MGG_07986.5were set up follow-ing the standard protocol of the Promega pGEM-T easy vector system.Transformation tests showed that ligation of the T-vector with the MGG_07986.5frag-ments yielded a large number of colonies,whereas ligation of the T-vector alone yielded only a few colonies (Fig.1B).Restriction digestion screening con-ﬁrmed that the plasmids yielded from ligation of theT-vector with the PCR product were true recombinants (Fig.1C).To establish a general guide for consistently successful cloning,several factors,such as Xcm I over-digestion for generating a T-vector,insert-to-vector molar ratios,and different T4DNA ligases,were tested to determine their effect on cloning efﬁciency.Surprisingly,we observed that T4DNA ligases could have a signiﬁcant impact on cloning efﬁciency.Liga-tions using Promega T4DNA ligase,the same product used by the pGEM-T easy vector system,consistently gave very high cloning efﬁciency with almost no self-ligation background.However,regular T4DNA li-gases from USB Corporation usually gave very low ligation efﬁciency for this TA cloning system (Fig.1B).Although the ligation efﬁciencies were a little higher at insert-to-vector molar ratios of 4:1to 8:1with the T-vector generated by standard digestion,ligations from vectors with 10-or 20-fold overdigestion and ligations with insert-to-vector molar ratios of 1:1,4:1,8:1,and 12:1all yielded good cloning efﬁciency when Promega T4DNA ligase was used (data not shown).Set of Expression ZeBaTA Vectors for PlantsUsing ZeBaTA,we developed a set of transient and stable expression vectors for different applications in both dicot and monocot plants.The backbone ofallFigure 2.Site-speciﬁc mutagenesis of the maize ubiqutin-1promoter (A)and the backbone of the binary vector pCAMBIA1300(B)in which three Xcm I recognition sites were deleted.The nucleotides represented in lowercase italic letters are the positions where deletions or mutations were made.Kan,Kanamycin resistance gene;LB,T-DNA left border;RB,T-DNA right border.C,Comparison of the levels of GUS expression mediated by the original and modiﬁed maize ubiquitin-1promoter in transiently transfected rice protoplasts.GUS activities are represented as a ratio of relative GUS/LUC.The experiment was repeated three times with similar results.1,Protoplast sample transfected with pUbiGUS;2,protoplast sample transfected with pXUN-GUS.A Zero Background Vector Systemtransient expression vectors is derived from pBlue-script II KS (),a high-copy-number cloning vector that can facilitate the isolation of a large amount of plasmid DNA for transient ex-pression.The backbone of all stable expression vectors is derived from pCAMBIA1300(),an Agrobacterium tumefaciens binary vector widely used for transformation in both dicot and monocot plants.Two different promoters,a cauliﬂower mosaic virus 35S promoter (Odell et al.,1985)and a maize (Zea mays )ubiquitin-1promoter (Christensen et al.,1992)were used to drive expression of genes of interest in dicots and monocots,respectively.The 35S promoter is more efﬁcient in dicots,whereas the maizeubiquitin-1Figure 3.ZeBaTA-based expression vectors for gene overexpression/silencing,protein tagging,protein subcellular localization,and promoter analysis in plants.A,Schematic structures of the transient expression vectors generated by Xcm I digestion.B,Schematic structures of the Agrobacterium -mediated stable transformation vectors generated by Xcm I digestion.LB,T-DNA left border;RB,T-DNA right border.Chen et al.promoter is more efﬁcient in monocots(Christensen et al.,1992).The original maize ubiquitin-1promoter and pCAMBIA1300vector,however,contain one and three Xcm I recognition sites,respectively(Fig.2,A and B).To facilitate the construction of the ZeBaTA-based expression vectors,the Xcm I recognition sites of the maize ubiquitin-1promoter and pCAMBIA1300vec-tor were eliminated by site-speciﬁc deletion or site-speciﬁc mutation(Fig.2,A and C).The designed expression vectors were all engineered with the cas-sette of the Xcm I-ccdB-Xcm I fragment(Supplemental Fig.S1).Figure3,A and B,illustrates the structural maps of the12transient and12stable transformation T-vectors.All vectors have been tested for cloning at least one time,and the results showed that these ZeBaTA expression vectors,including those binary vectors that are relatively large in size(.10kb), consistently yielded high cloning efﬁciency(Supple-mental Fig.S2).Because the maize ubiquitin-1promoter used in this system was modiﬁed to block its original Xcm I recog-nition site by deleting a single base(Fig.2A)at the position of nucleotide2480,a gus gene(Jefferson et al., 1987)was ampliﬁed by PCR and then cloned into the Xcm I-digested pXUN vector to produce an expression construct to test the expression activity of the modiﬁed maize ubiquitin-1promoter.The derived constructpXUN-GUS and a control construct pUbi-GUS(Chen et al.,2006b),of which a gus gene is driven by the original maize ubiquitin-1promoter,were tested tran-siently in the transfected rice protoplasts.Transient expression assays showed that the levels of GUS activity in rice protoplasts transfected with these two constructs were similar(Fig.2C),indicating that the deletion of nucleotide2480does not affect the activity of the maize ubiquitin-1promoter.Testing of Tagged Protein ExpressionEpitope tagging is a widely used method for the rapid and effective characterization,puriﬁcation,and in vivo localization of the protein products of cloned genes.To facilitate gene cloning for epitope tagging in plants,a total of12epitope-tagging vectors(Fig.3,A and B)were constructed using ZeBaTA.These vectors contain a35S promoter or a maize ubiquitin-1pro-moter,allowing direct cloning of genes of interest into expression vectors to express a translational fusion of target protein with three commonly used epitope tags in plants(i.e.FLAG,HA,or Myc;Earley et al.,2006). To determine the feasibility of this epitope-tagging system,a gfp gene was cloned into the pXUN-HA vector to fuse with the HA tag.The resulting expres-sion construct pXUN-HA-GFP was transiently ex-pressed in rice protoplasts.As shown in Figure4,A and B,protoplasts transfected with pXUN-HA-GFP showed strong GFPﬂuorescence,and HA-tagged GFP protein was detected in protein extracts of trans-fected protoplasts but not in the nontransfected con-trol,demonstrating the potential application of this system for functional study of target proteins in plants.Gene Silencing by Hairpin RNAi or Artiﬁcial MicroRNA In plants,a typical and efﬁcient approach to induce gene silencing is to use an inverted-repeat construct to express hairpin RNA(hpRNA;Waterhouse et al.,1998; Smith et al.,2000).However,a major limitation of the hpRNA interference(hpRNAi)approach for high-throughput gene functional analysis is the cumber-some cloning procedure for generating hpRNAi constructs(Helliwell and Waterhouse,2003).The gen-eration of a hpRNAi construct using conventional restriction enzyme digestion and DNA ligation meth-ods usually requires several cloning steps.Although Gateway cloning technology has been adapted to gen-erate hpRNAi constructs(Helliwell and Waterhouse, 2003;Miki and Shimamoto,2004),it still requires two cloning steps.With the ZeBaTA system,hpRNAi con-structs can be made by a single-step cloning procedure (Fig.5A).Instead of making an inverted-repeat cas-sette by DNA recombination techniques,we designed a new approach to assemble the hpRNAi cassette by overlapping PCR.Brieﬂy,a target fragment with an additional3#-terminal sequence complementary to both the5#-and3#-terminal ends of a designed spacer fragment is ampliﬁed as aﬁrst step.The overlapping fragments are then fused together in a subsequent PCR reaction,and the resulting inverted-repeat is cloned directly into a ZeBaTA expression vector(Fig.5A).To test the feasibility of this approach,an RNAiconstruct Figure4.Transient expression and protein-tagging detection of the ZeBaTA vectors in rice protoplasts.A,Fluorescence microscopy of the expression of HA-tagged GFP in rice protoplasts.B,Detection of HA-tagged GFP by western ne1,Nontransfected control protoplast sample;lanes2to4,independent protoplast samples transfected with pXUN-HA-GFP.[See online article for color version of thisﬁgure.]A Zero Background Vector Systemwas generated by overlapping PCR in which the sense and antisense 217-bp fragments of the Arabidopsis phytoene desaturase gene (PDS )were separated by a 420-bp stuffer fragment derived from the gus gene.The resulting fragment was cloned into the pCXSN vector (Fig.3B)to generate the expression construct pCXSN-atPDS-RNAi.The RNAi construct was introduced into Arabidopsis by the ﬂoral-dip method.Over 80%of transgenic plants had a clear albino phenotype (Fig.6A),a typical visible phenotype caused by silencing of the PDS gene (Guo et al.,2003;Miki and Shimamoto,2004).Recently,the artiﬁcial microRNA (amiRNA)ap-proach has been introduced for highly speciﬁc gene silencing in both dicot and monocot plants (Niu et al.,2006;Schwab et al.,2006;Ossowski et al.,2008;Warthmann et al.,2008).Typically,the amiRNA is generated by site-directed mutagenesis on precursors of endogenous miRNAs to exchange the natural miRNA sequences with those of amiRNAs using overlapping PCR (Ossowski et al.,2008).The same ZeBaTA-based vector system developed for ectopic gene expression and hpRNAi can also be used for making amiRNA expression constructs by simpleTAFigure 5.Schematic illustration of the construction of hpRNAi or amiRNA constructs by single-step cloning.A,Generation of hpRNAi constructs by overlapping PCR approach.The target gene fragment and the stuffer sequence fragment are ampliﬁed in the ﬁrst-round PCR.Primers P2,P3,and P4introduce complementary adapters (indicated by vertically lined boxes)to the ampliﬁed fragments.The two ampliﬁed fragments are fused together as an inverted-repeat cassette in the second-round PCR by using single P1primer.The resulting fragment is then directly cloned into the plant expression T-vector.B,Generation of amiRNA constructs by overlapping PCR approach.C,Generation of amiRNA constructs for rice genes by single-step PCR.The expression vectors pXUN-osaMIR528and pCXUN-osaMIR528were preassembled with 5#and 3#stemloop backbone sequences of a rice miRNA precursor Osa-MIR-528(Warthmann et al.,2008).Thus,making amiRNA constructs for rice target genes only requires an amiRNA-amiRNA*fragment generated from single-step PCR.The nucleotides represented in lowercase letters are the positions where mutations were made to introduce two Xcm I recognition sites.Chen et al.cloning (Fig.5B),thus bypassing the time-consuming two-step procedure for the regular restriction enzyme digestion-mediated cloning or the Gateway cloning (Ossowski et al.,2008).We further developed a ZeBaTA-amiRNA system to simplify the generation of rice amiRNA constructs because our lab is focusing on rice functional genomics.The new ZeBaTA-amiRNA vector was designed based on the stemloop backbone derived from Osa-MIR528,an endogenous rice miRNA precursor that has been used to efﬁciently express amiRNAs for highly speciﬁc silencing of targeted genes in rice (Warthmann et al.,2008).By site-directed muta-genesis of a single base on the 5#and 3#stemloop backbones of Osa-MIR528,respectively,a cassette of 5#Osa-MIR528stemloop backbone-Xcm I-ccdB -Xcm I-3#Osa-MIR528was assembled and cloned into the expres-sion vectors where the expression of amiRNA is under the control of the maize ubiquitin-1promoter.Figure 5C illustrates the structural maps of the Osa-MIR528-based vectors pXUN-osaMIR528and pCXUN-osaMIR528.The vectors allow for high-throughput generationof rice amiRNA constructs by cloning the amiRNA-amiRNA*fragment generated from a single-step PCR into the ZeBaTA vector with the preassembled Osa-MIR528stemloop backbone (Fig.5C;Supplemental Fig.S3),thus avoiding the time-consuming overlapping PCR.The modiﬁed vector was evaluated by expression of the amiRNA for silencing of the OsPDS gene.The two constructs pCXUN-amiPDS and pCXUN528-PDS,which contain original or modiﬁed Osa-MIR528stem-loop backbone with amiRNA sequence targeting OsPDS ,respectively ,were introduced into rice cv Nip-ponbare by Agrobacterium -mediated transformation.Consistent with a previous study (Warthmann et al.,2008),70.1%of the primary transgenic lines transformed with pCXUN-amiPDS had a bleaching PDS silencing phenotype (Fig.6B;Table I).Similarly,77.1%of the primary transgenic lines transformed with pCXUN528-PDS had the same albino phenotype,suggesting that the mutagenesis on the Osa-MIR528stemloop backbone does not affect the biogenesis of the amiRNA for silenc-ing of the PDS gene.Protein Subcellular Localization/Colocalization and Promoter Activity AssayTo investigate the subcellular localization or colo-calization of particular proteins,a set of ZeBaTA vectors (i.e.pXDG,pXDR,pCXDG,and pCXDR)was devised for transient or stable expression of protein fusions with GFP or red ﬂuorescent protein.The vectors contain a 35S promoter-driven gfp or DsRed cassette that has been used to visualize protein local-ization in both dicot and monocot plants (Goodin et al.,2002;Chen et al.,2006b).As shown in Figure 3,A and B,PCR products of genes of interest can be simply engineered into the vectors to fuse with the gfp or DsRed gene.To conﬁrm whether the vectors can be used for detecting protein localization in plant cells,the rice Spin1gene encoding a putative RNA-binding protein previously shown to be nuclear targeted(Vega-Sa´nchez et al.,2008)was cloned into vectors pXDG and pXDR to fuse in-frame with gfp and DsRed ,respectively.Transient expression of the constructs pXDG-Spin1and pXDR-Spin1in rice protoplasts dem-onstrated that the GFP-and DsRed-SPIN1fusion proteins were targeted to the nuclear region as pre-dicted (Fig.7A).For promoter activity assays,two reporters,gus and gfp ,were used for constructing pXGUS-P/pCXGUS-P and pXGFP-P/pCXGFP-P ,respectively.The linear T-vectors of pXGUS-P/pCXGUS-P orpXGFP-P/Figure 6.Silencing of the PDS gene in Arabidopsis and rice by the ZeBaTA-based hpRNAi or amiRNA approaches.A,Arabidopsis plants transformed with the hpRNAi construct pCXSN-atPDS-RNAi showing the PDS silencing albino phenotype.(1)Control plant;(2and 3)two examples of transgenic Arabidopsis plants.B,Rice plants transformed with the amiRNA vectors showing the albino phenotype.(1)Control plant;(2)example of pCXUN-amiPDS-transformed plants;and (3)example of pCXUN528-PDS-transformed plants.C,RT-PCR analysis of PDS suppression transgenic rice plants.Five independent primary plants (1,2,3,4,and 5)transformed with pCXUN-amiPDS and ﬁve independent primary plants (6,7,8,9,and 10)transformed with pCXUN528-PDS were selected for the analysis.CK,Wild-type Nip-ponbare plant used as the control.Table I.PDS silencing frequency of transgenic rice mediated by the ZeBaTA-amiRNA systemamiRNA VectorTotal Independent TransformantsAlbino PhenotypeEfﬁciency%pCXUN-amiPDS 553970.1pCXUN528-PDS 352777.1A Zero Background Vector SystempCXGFP-P (Fig.3,A and B)allow direct cloning of PCR-ampliﬁed promoter fragments located in front of the reporter genes.As proof of concept,the 35S pro-moter was cloned into pCXGUS-P to drive expression of the reporter gene gus .Arabidopsis plants stably transformed with the construct pCX-35S-GUS showed constitutive GUS expression in the whole plants (Fig.7B),conﬁrming the feasibility of the system for assay-ing promoter activity.DISCUSSIONWith the rapid development of the next-generation sequencing technology,more plant genomes will be sequenced in the near future.How to rapidly deter-mine the function of the identiﬁed genes on a large scale is a daunting challenge.The ability to efﬁciently make constructs to transiently and stably express speciﬁc genes in cells,tissues,or whole plants is a fundamental aspect and bottle neck of plant functional genomics research.Traditionally,the cloning vectorsfor plant research carry a multicloning site (MCS)within their target gene expression cassettes.The restriction sites in the MCS are rather limited,making cloning of most target genes difﬁcult.Although TA cloning vectors have been widely used for cloning of PCR-ampliﬁed fragments,the system has not yet been incorporated in the cloning vectors for transient and stable expression of target genes because of the tech-nical challenge of generating low-background TA cloning vectors.The Gateway system has been a popular choice for generating various constructs be-cause it allows the gene of interest to be easily cloned into speciﬁcally designed plasmids without DNA re-striction digestions.The two-step cloning and expen-sive reagents,however,make the Gateway system impractical for large-scale cloning in most individual laboratories when the entry clone collections are not available.The ZeBaTA system described here over-comes the limitations of both the TA and Gateway cloning systems.After two Xcm I recognition sites have been introduced into the MCS,any PCR fragments with a T-overhang can be easily cloned into a ZeBaTA vector.With the introduction of the negative selection marker gene ccdB between the two Xcm I sites,any self-ligation transformants are ing this tech-nology,we constructed a set of 12transient and 12stable transformation vectors for plant gene expres-sion studies and tested the vectors in rice or Arabi-dopsis in our laboratories.These vectors can be used in a wide range of functional genomics projects in plants and will be distributed to the research community upon request.Under certain conditions,cloning with T-vectors generated by digestion with Ahd I or Xcm I gave low efﬁciency and the T residue of the insert-vector junc-tion in the recombinant clones is often missing (Mead et al.,1991;Chen et al.,2006a).Chen et al.(2006a)speculated that this may be due to the presence of unknown factors that,during digestion and prepara-tion of the T-vectors,inﬂuence the stability of 3#-T overhangs.In this study,we found that the main factor affecting successful cloning is the use of an appropri-ate T4DNA ligase.We tested the Promega T4DNA ligase,which is included in the pGEM-T easy vector system,and the T4DNA ligase from USB Corporation.The ligations using Promega T4DNA ligase consis-tently gave a very high cloning efﬁciency;most of the ligations using USB Corporation T4DNA ligases yielded low efﬁciency.Many of the recombinant plas-mids from the latter ligations missed a T residue in the insert-vector junction,consistent with observations by Mead et al.(1991)and Chen et al.(2006a).The T residue is missing mainly because regular commercial T4DNA ligases contain exonuclease activities that can remove the 3#-T tails from the vector,as reported in the technical manual of the pGEM-T and pGEM-T easy vector systems (/tbs/tm042/tm042.pdf);removal of the 3#-T tails from the vector results in very low cloning efﬁciency.When the Promega T4DNA ligase was used for ligation,weFigure 7.Protein subcellular localization and promoter activity anal-ysis using the ZeBaTA vectors.A,Fluorescence microscopy of the coexpression of GFP and DsRed,or GFP-SPIN1and DsRed-SPIN1fusions in rice protoplasts.Scale bar =20m m.The RNA binding nuclearprotein SPIN1was used as a tester (Vega-Sa´nchez et al.,2008).B,GUS staining of Arabidopsis transformed with pCX-35S-GUS,where the 35S promoter was cloned into the vector pCXGUS-P to test the system.CK,Plant transformed with control vector pCAMBIA1300();pCX-35S-GUS-1and pCX-35S-GUS-2,two independent primary transgenic plants.Chen et al.。

IBM Z Batch Resiliency V1R1 用户指南和参考说明书

Preface............................................................................................................... xxi
Who should read this book........................................................................................................................xxi What this book contains............................................................................................................................ xxi Publications............................................................................................................................................... xxi Accessibility............................................................................................................................................... xxi Support information.................................................................................................................................. xxi

Eclipse数值模拟软件问答

Eclipse数值模拟软件问答（初级）1. ECLIPSE输出结果文件是哪些？.GRID或.FGRID: 网格文件.EGRID: 网格文件，与GRID格式不同，文件要小的多。

（用关键字GRIDFILE来控制输出类型）.INIT或.FINIT: 属性文件。

（用关键字INIT来控制输出）.PRT: 报告输出。

文件很大，模型处理及计算结果详细报告。

（RPTGRID，RPTPROP，RPTSOL，RPTSCHED 控制输出）.LOG: 后台作业时的输出报告，文件比PRT要小很多。

可用于错误检查。

.DBG: Debug文件，一般不用。

可用于检查ECLIPSE如何处理输入参数。

.SAVE: 用于快速重启。

（用关键字SAVE来控制输出）.RFT：RFT计算结果。

（用关键字WRFTPLT来控制输出).FLUX: 流动边界。

(用关键字DUMPFLUX来控制输出).Snnnn或.UNSMRY: 图形文件输出（在SUMMARY部分定义）.Xnnnn或.UNRST: 重启文件输出（用RPTRST，RPTSOL或RPTSCHED来控制输出）2. ECLIPSE输出文件都有什么格式？格式化输出：可读文件，文件大。

（用关键字FMTOUT来控制）非格式化输出：不可读文件，文件小。

多输出文件：每一时间步一个输出文件。

单文件输出：所有时间步输出到一个文件。

（用关键字UNIFOUT来控制)ECLIPSE缺省输出：非格式化，多文件输出。

3. ECLIPSE数据文件分几部分，各部分定义什么数据类型？ECLIPSE数据类型分八部分，各部分内的关键字除几个个别的外不能混用。

RUNSPEC: 定义模型维数以及模型基本类型，包括模型网格维数，最大井数，井组数，流体类型，输出类型控制等。

GRID: 定义模型网格和属性，包括顶部深度，厚度，孔隙度，渗透率，净毛比，一般由前处理软件Flogrid或Petrel输出。

EDIT: 编辑孔隙体积，传导率。

Schneider-Electric IH 24h 时间开关说明书

D 15365-561-03_e n 12/2020¼DANGERRisk of serious damage to property and per-sonal injury, e.g. from fire or electric shock, due to incorrect electrical installation.Safe electrical installation can only be ensured if the person in question can prove basic knowledge in the following areas:•Connecting to installation networks •Connecting several electrical devices •Laying electric cablesThese skills and experience are normally only pos-sessed by skilled professionals who are trained in the field of electrical installation technology. If these minimum requirements are not met or are disregard-ed in any way, you will be solely liable for any dam-age to property or personal injury.IH 24h is a mechanical time switch that switches con-nected loads on or off when the set time has been reached. It is installed on a DIN rail (DIN EN 60715).1Place the IH 24h onto the DIN rail.2Connect cables:–Remove 8 mm (max. 9 mm) of insulation –Open the plug-in terminal with a screwdriver and plug in the cable at a 45° angle. (max. 2 cables per plug-in terminal)IH 24hOperating instructionssrm: CCT16364 arm: CCT15365For your safetyG etting to know IH 24hProduct detailsA Manual switchB Setting discC SwitchingsegmentsD Automatic/perma-nent switchE Clock hands (hour, minutes)F Display: Morning(3/6/9), afternoon (15/18/21)G Rotary knob H Switch output I Mains connectionInstalling IH 24h3Connect the mains voltage.|IH 24h arm (art. no. CCT15365) is equipped with a quartz clock mechanism. The quartz clock mechanism only starts after a few minutes after having connected it a power supply. Complete power reserve is reached after 5 days.Setting the timeUse the rotary knob to set the time (hour, minutes). Y ou can turn the rotary knob clockwise and anti-clockwise.|The clock face changes between morning and af-ternoon times when the dial has gone past 24 and 12 hours.Setting the switching timeY ou can set the switching time using the switching seg-ments. Each switching segment stands for a 15-minute time period. The switching segments can be pushed in or out, for example, by using your index finger. The set-ting disc shows the switching period (+/- 5 minutes).Operating the manual switchY1Turn the manual switch by one position anti-clockwise.The current status is reversed and stays unchanged forSetting IH 24hTo set the time in the morning, 3/6/9 must be visible on the clock face.To set the time in the after-noon, 15/18/21 must be visible on the clock face.Switching segment out Load switched off Switching segment inLoad switched on Switching time 1 = 6:00 - 10:00Switching time 2= 17:00 - 21:00Operating IH 24hOperating the automatic/permanent switch With the automatic/permanent switch, you can switch the load permanently on or off or permanently set it to au-tomatic mode.•Set the switch to "1".The load is switched on permanently. The set switch-ing times are deactivated.•Set the switch to "0".The load is switched off permanently. The set switch-ing times are deactivated.•main activated. The load is switched on or off when the set switching time has been reached.|For permanent ON or permanent OFF , the set switching times are deactivated.If you have technical questions, please contact the Customer Care Centre in your country. T echnical dataNominal voltage:CCT16364AC 230 V , 50/ Hz CCT15365AC 110-230 V , 50-60 Hz Nominal current:16 A, cos φ = 14 A, cos φ = 0.6Incandescent lamps:AC 230V , max. 1100W Halogen lamps:AC 230V , max. 1000W Fluorescent tubes:AC 230V , max. 600VA Fluorescent lamps with electronic ballast:2 x 40W (12µF), parallel-compensatedCompact fluorescent lamps with electronic ballast:25 W LED lamps:<2W: 20 W>2W:180 W Power consumption:≈ 0.5 W Ambient temperature:-20°C to +55°C Connecting terminals: 2 x 0.5 - max. 2.5 mm², fixed and flexible wires Power reserve:CCT15365150 h (230V), 75 h (110V)Accuracy:CCT16364Network synchronous CCT15365≤ ±1s/day at +20°C Mode of operation:srm: Device of 1BRTU type in accordance with EN 60730-1arm: Device of 1BSTU type in accordance with EN 60730-1Degree of pollution:2Rated impulse voltage:4000 VProtection class:II per EN 60730-1 when installed correctly T ype of protection:IP 20 in accordance with EN 60529Schneider Electric Industries SAS。

MVTS基本使用说明

MVTS安装使用说明（适用310 312）第一节，安装操作系统要求：310 运行在LINUX AS3或ES3 以下的版本312运行在LINUX Radhat 9.0以上版本，（不包括LINUX9.0）建议硬件配置：CPU：P4 2.0以上内存：2G以上硬盘：30G以上安装步骤：安装linux完成后，将MVTS的manager.tar.gz和MVTS-310-Linux.tar.gz两个安装包上传到/usr/local 目录下.使用SSH登陆LINUX，执行以下命令：cd /usr/loca 回车tar -zxvf MVTS-310-Linux.tar.gz 回车tar –zxvf manager.tar.gz 回车cd mvts 回车[root@localhost mvts]# sh setup.shMVTS installation:enter MVTS admin group id:0 输入0回车enter MVTS support group id:0 输入0回车enter MVTS billing group id:0 输入0回车updating ./cfg/meraproxy.cfg ...setting permissions to MVTS contents ...updating /etc/profile ...making startup script mvts ...moving startup script to /etc/rc.d/init.d ...adding links for start/stopcreate links:/etc/rc.d/init.d/mvts <- /etc/rc.d/rc5.d/S50mvts/etc/rc.d/init.d/mvts <- /etc/rc.d/rc0.d/K50mvtsinstallation successful这样MVTS就安装好了，这个时候还不能登陆，需要安装managercd ..回车cd manager 回车sh setupMVTS directory? [/usr/local/mvts]: /root/mvtsMVTS Manager directory? [/root/mvts/manager]: 回车MVTS admin group? [mvts]: 回车Group mvts does not exist. Create? [y/n]: yMVTS admin user (not root)? [mvts]: 回车User mvts does not exist. Create? [y/n]: yChanging password for user mvts.New password: 密码写入linux的密码BAD PASSWORD: it is too shortRetype new password: 密码写入linux的密码passwd: all authentication tokens updated successfully.MVTS Manager successfully installed.Please do following steps before using MVTS Manager:1. Edit file /root/mvts/cfg/meraproxy.cfg:[Console]console_port=1730admin_gid=5012. Reload MVTS configuration:/root/mvts/bin/mp_shell.x r c -d3. Start MVTS agent from root:/root/mvts/manager/bin/mvtsagntctl startor/etc/init.d/mvtsagnt startThank you for using MVTS Manager![root@localhost manager]# /etc/init.d/mvtsagnt startStarting MVTS Agent: [ OK ]Manager 安装最好装两次，也就是执行2次sh setup 确保能安装成功请注意以上的红色，那写请按上面的中文提示作相应操作，安装完成后先别急着启动请注意这段提示：1. Edit file /root/mvts/cfg/meraproxy.cfg:[Console]console_port=1730admin_gid=501这里也要做相应修改写入vi /root/mvts/cfg/meraproxy.cfg回车找到后面这个按A作相应修改：console_port=1730admin_gid=501修改好后按ESC退出编辑命令，再输入wq! 保存退出以下命令是用来启动和停止MVTS的起动MVTS: /etc/init.d/mvts start停止MVTS: /etc/init.d/mvts stop起动mvtsagnt: /etc/init.d/mvtsagnt start停止mvtsagnt: /etc/init.d/mvtsagnt stop安装完成后第二节：MVTS的登陆和相关功能按键介绍请在你的电脑上安装MVTS的管理软件MVTSmanager并运行manager软件看到以上窗口说明MVTSmanager成功运行了，这个时候我们要做登陆服务器的数据:点右边的Create 看到如下窗口:写入相关数据，MVTS刚安装的默认登陆端口是1730 帐号和密码是admin 都是小写，小提示:MVTS的数据里面不可以存在全角字符更不能有中文，不然是不能登陆的.写完后点OK就保存下来了，然后选中他双击或点右上解的OK，即可连接，连接的过程会有点慢，请耐心等待，出现如下窗口后即说明成功登陆了:本文重点介绍如何对接IP认证客户和、注册用户、和落地对接路由方式←落地对接和网关对接都是在这里←路由调度在这里←注册帐号类型Originators 是指用户的帐号品质报告Terminators 是指落地的品质报告Gteways 是指所有在Gateways的品质报告Dialpeers是指路由调度的品质报告Endpoints 这里显示RAS 注册帐号的在线状态Active Calls是指当前在线的活动呼叫CDR 显示最近100条目的是用来查线路的问题Edit Config 编辑模式，要作数据改动，必须进入此模式（同时只能一人在线）Cancel edit 撤消编辑并退出。

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3
Figures Of Unity
1. Pleasant And Sweet as Aaron’s Perfumed Oil
– High Priest Presented One Holy Nation – United In Worship To The One God
2. Refreshing And Invigorating As Morning Dew
9
Five Step Walk For Unity Ephesians 4:1-3
• • • • • Walk Worthy Of Our Calling Walk With All Lowliness And Meekness Walk With Longsuffering Walk Forbearing One Another In Love Walk Endeavoring To Keep The Unity Of The Spirit In The Bond Of Peace
14
Al Maxey’s Unity Eliminate Doctrine (Pattern)
• Reflections – Al Maxey Issue #3 December 12, 2003
• “When God provided His people with guiding principles did He honestly expect all peoples everywhere until the end of time to implement those principles ONLY after the manner of the "approved examples" of a band of fallible disciples in one small corner of the globe? And if so, where has God ever declared this precept? Where has God declared specifically that unless we give or love or pray or sing JUST EXACTLY LIKE they did in Troas or Corinth or Ephesus or Jerusalem we will go straight to hell and be tortured for a zillion years?! Would someone please show me where God ever declared such a thing? What specific command of God makes the methodologies of these early disciples the universal standard for all peoples for all time?! I have yet to find that clear command in Scripture.ers in open rebellion and sin
Text For Our Study
Ephesians 4:1-6 • I therefore, the prisoner in the Lord, beseech you to walk worthily of the calling wherewith ye were called, 2 with all lowliness and meekness, with longsuffering, forbearing one another in love; 3 giving diligence to keep the unity of the Spirit in the bond of peace. • 4 There is one body, and one Spirit, even as also ye were called in one hope of your calling; 5 one Lord, one faith, one baptism, 6 one God and Father of all, who is over all, and through all, and in all.
– – – –
Accepting them in their sin Doctrine takes backseat for sake of uniting Brethren determine to agree to disagree Doctrinal disagreement is accepted as the norm
10
Man’s Part – Five Steps To Unity
1. Walk Worthy Of Our Calling
– Walk
• • • • • • • • • In Newness of life – Rom. 6:4 Not after flesh but the spirit – Rom. 7:14-25 Have the disposition to follow God – Rom. 8:4 As children of light – Eph. 5:8 By faith not by sight – 2 Cor. 5:7 Let us walk by the same rule – Phil 3:16 Called by the Gospel – 2 Thess 2:14-15 Therefore we must stand fast in that Gospel Hold Fast the traditions taught : word or epistle
13
Leroy Garrett Restoration Review Vol. 20, No. 10; Dec. 1978
• “It is love that unites, not doctrinal agreement. Love perfectly unites that which is divided. Even if people should reach perfect agreement on all the points of doctrine, this would not mean perfect unity. Only love made for perfect unity, and this when folk may be quite diverse in their interpretation of much of the Bible.”
• • • • • Al Maxey rejects the scriptures being a pattern to emulate Rubel Shelly and Max Lucado accept sinners prayer Ketcherside and Garrett accepts non-Christians Brothers in prospect becomes brother in fact Others accept same principle in a limited fashion
• • • • • Paul tells us unity is possible That God demands we have it We must endeavor to maintain it Common mistake to skip our part Immediately Focus on God’s platform
Al Maxey’s Unity Remove Us From The Primitive Model
• Reflections –Al Maxey Issue #3 December 12, 2003
• “I personally doubt ANY of the early disciples would recognize much of anything we do today, especially in our worship assemblies on Sunday morning. In external FORM we are so far removed from anything found among first century disciples that they would likely disfellowship the whole bunch of us (assuming, of course, that they were equally into this whole patternistic, exclusivistic, isolationist nonsense ... which I do not for an instant believe they were).”
Unity Of The Spirit
In The Bond Of Peace
1
1. Introduction To Unity
2
Psalm 133
• Behold, how good and how pleasant it is For brethren to dwell together in unity! 2 It is like the precious oil upon the head, That ran down upon the beard, Even Aaron’s beard; That came down upon the skirt of his garments; 3 Like the dew of Hermon, That cometh down upon the mountains of Zion: For there Jehovah commanded the blessing, Even life for evermore.