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阿尔法拉維ATEX加权系統UltraPure輕芯说明书

The Alfa Laval weighing system is a complete solution offered for process weighing installation,where level measurement,mixing,ﬁlling,dosing or batching is required.The weighing solution is as standard delivered in three different accuracy ranges:0.10%,0.05% and0.025%with a total measuring range from0to4000kg.Each weighing system consists of1to4load cells and a weighing module. The weighing modules are available with both analog4-20mA output andﬁeldbus interface(PROFINET,Proﬁbus DP or EtherNet IP).For high hygienic demands,the Alfa Laval load cells are supplied electropolished and hermetically sealed to IP68(laser welded).Capacitive measurement principle(patented)The Alfa Laval robust digital load cells are based on a patented capacitive measurement principle where a non-contacting capacitive sensor is mounted inside the load cell body.As the capacitive sensor is not in contact with the load cell body,the load cells are to a very high degree unaffected by overloads,sideloads,torsion and welding voltages.Therefore,a straightforward and hygienic mechanical installation of the load cells can be done without expensive and complicated mounting kits and overload protection devices.The electrical installation of the digital load cells is pure plug-and-play as the signal from the non-contacting capacitive sensor is directly converted,compensated and calibrated by a patented ASIC.The digital signal is transmitted as RS485data on a reliable RG-58single wire coaxial cable which may be up to50meters long.The factory calibration of the digital load cells is independent of the load cell cable length.Technical dataStainless steel enclsure..........IP68Stainless steel enclosure with panelmounted display...............IP64Measuring range:..............from0to4000kg dependingon system selection. Accuracy:...................0.10%,0.05%,0.025% Compensated temperature range:...-10to50°COverload and sideload:..........300%overload tolerance Power supply:................24VDC,±10%min.2A Certiﬁcates•CE marked•Calibration certiﬁcate(option)• 3.1B certiﬁcate(option)Mechanical dataWeight:BL-EX Load cell:.............. 2.3kgWeighing module:..............approx.0.5kgMaterials:Load cells:..................AISI316and17-4PH Mounting kit:.................AISI304/AISI316(weldingcylinder)Operating temperature range:BL-EX Load cells:..............-40to50°CWeighing modules:.............-10to50°CProtection class:Load cells:..................IP68Weighing modules:.............IP20SpeciﬁcationParameter Unit0.10%0.05%0.025% Rated capacity(Emax)per load cell kg10,20,50,100,150,250,500,1000 Safe overload limit%of E max300to1000Safe sideload limit%of E max500to2000Minimum dead load%of E max0Accuracy%of E max0.1000.0500.025 Repeatability%of E max0.0180.0150.010 Hysteresis%of E max0.0330.0200.017 Creep30min.%of E max0.0350.0250.017 Temperature effect on zero%/10°C0.0400.0300.016 Temperature effect on sensitivity%/10°C0.0400.0300.016 Deﬂection at Emax mm max.0.10Mesuring rate Hz up to1000Internal resolution Bit24Maximum cable length m100OptionsOUTPUT:4-20mAPROFINETEtherNet IPProﬁbus DPRS485Local weighing display:Alfa Laval weighing terminal(options)Load cell cable:6m ATEX coaxial RG58with BNC connector(option:10,20or50m)Mounting:Mounting kit for BL-EX beam load cellDimensional drawingsBL-EX Beam load cell0-100kg with mounting kitBL-EX Beam load cell100-1000kg with mounting kitLayout and electrical connection schematic of weighing modules:Proﬁnet weighing module for4load cells(4XXXA)(external dimensions is the same for Proﬁlbus DP and Ethernet IP4-20mA output weighing module for4load cells(4X79A)RS485interface module(when ordered with display(4X40A)ATEX POWER SUPPLY(4051A)14mm/5.51in128mm/5.4in116mm/4.57in11mm/.43in29 mm / 1.14 in60 mm / 2.36 inø4.5 mm / ø0.18 in66 mm / 2.60 in14mm/5.51in128mm/5.4in116mm/4.57in11mm/.43in29 mm / 1.14 in60 mm / 2.36 inø4.5 mm / ø0.18 in66 mm / 2.60 inInstallation in ATEX zonesLoad cells x=number of loadcellsATEX Power Supply Gnd-+Interfaces: EtherNet IPPROFINET EtherCAT Proﬁbus DP Modbus TCP/IP DeviceNet RS485Power Supply +24Vdc,2AHAZARDOUS AREA SAFE AREASelection guideWhen conﬁguring a weighing system,you need the following information:•ATEX application•Number of tank legs or supporting points•Total weight of tank incl.product in kg•Required output and/or local display•Required accuracy for the application(e.g.dosing,mixing,level measurement etc.)With this information,you are able toﬁnd the conﬁguration you need in the pricelist or in the online conﬁguration tool:Step1:Is the weighing system used in ATEX zone1,2,21,22classiﬁed operation.Step2:Calculate the total weight of the tank inclusive the product in kg and round up to the nearest standard load cell system.Step3:Decide on accuracy required by the application-0.10%accuracy systems are suitable for mixing applications-0.05%accuracy systems are suitable for dosing applications-0.025%accuracy systems are suitable for very precise dosing and batching applicationsStep4:Decide on the output signal type and/or a local weighing display:-4-20mA-PROFINET-EtherNet IP-Proﬁbus DP Step5:Decide on the length of the load cell cables(the length of the cable is can be shortened without the need for recalibration)-6m(standard)-10m-20m-50mStep6:Decide on if you need calibration certiﬁcateTheoretical statistical weighing system accuracy System range System rangeItem no.3supporting points4supporting pointsNumber ofloadcells andlc type in systemSystem type0.10%System type0.05%System type0.025%TE67WB9KXXXXXX0-60kg.(132lb)(3*20kg.lc)0.035kg.(0.076lb)0,017kg(0.038lb)0.009kg.(0.019lb) TE67WBBJXXXXXX0-80kg.(176lb)(4*20kg.lc)0.040kg.(0.088lb)0,020kg.(0.044lb)0.010kg.(0.022lb) TE67WBCKXXXXXX0-90kg.(198lb)(3*30kg.lc)0.052kg.(0.115lb)0,026kg.(0.057lb)0.013kg.(0.029lb) TE67WBEXXXXXXX0-120kg.(265lb)(4*30kg.lc)0.060kg.(0.132lb)0.030kg.(0.066lb)0.015kg.(0.033lb) TE67WBFKXXXXXX0-150kg.(331lb)(3*50kg.lc)0.087kg.(0.191lb)0.043kg.(0.095lb)0.022kg.(0.048lb) TE67WBGKXXXXXX0-200kg.(441lb)(4*50kg.lc)0.100kg.(0.220lb)0.050kg.(0.110lb)0.025kg.(0.055lb) TE67WBIXXXXXXXX0-300kg.(661lb)(3*100kg.lc)0.173kg.(0.382lb)0.087kg.(0.191lb)0.043kg.(0.095lb) TE67WBJXXXXXXXX0-400kg.(882lb)(14*100kg.lc)0.200kg.(0.441lb)0.100kg.(0.220lb)0.050kg.(0.110lb) TE67WBNKXXXXXX0-450kg.(992lb)(3*150kg.lc)0.260kg.(0.573lb)0.130kg.(0.211lb)0,065kg.(0.143lb) TE67WBOXXXXXXX0-600kg.(1323lb)(4*150kg lc)0.300kg.(0.661lb)0.150kg.(0.331lb)0.075kg.(0.165lb) TE67WBPXXXXXXX0-750kg.(1653lb)(3*250kg lc)0.433kg.(0.955lb)0.217kg.(0.477lb)0.108kg.(0.239lb) TE67WBLXXXXXXX0-1000kg.(2205lb)(4*250kg lc)0.500kg.(1.102lb)0.250kg.(0.551lb)0.125kg.(0.276lb) TE67WBSXXXXXXX0-1500kg.(3307lb)(3*500kg.lc)0.866kg.(1.909lb)0.433kg.(0.955lb)0,217kg.(0.477lb) TE67WBTXXXXXXX0-2000kg.(4409lb)(4*500kg.lc) 1.000kg.(2.205lb)0.500kg.(1.102lb)0.250kg.(0.551lb) TE67WBUKXXXXXX0-3000kg.(6614lb)(3*1000kg.lc) 1.732kg.(3.819lb)0.866kg.(1.909lb)N/ATE67WBVXXXXXXX0-4000kg.(8818lb)(4*1000kg.lc) 2.000kg.(4,409lb) 1.000kg.(2.205lb)N/ANote:All calculations are the theoretical worst case accuracy that can be obtained with the Alfa laval weighing system solutions.The presented data It is solely for Informational purpose,all real life accuracies is highly dependable of proper weighing system installationAlfa Laval reserves the right to change specifications without prior notification.How to contact Alfa Laval Contact details for all countriesare continually updated on our website.Please visit to access the information direct.A l f a L a v a l i s a t r a d e m a r k r e g i s t e r e d a n d o w n e d b y A l f a L a v a l C o r p o r a t e AB . 100001495e n 1906。

细胞周期素A2检测试剂盒说明书

细胞周期索A2检测试剂盒说明书实验原理：本试剂盒应用双抗体夹心法测定标本中血清素/血清胺(ST)水平。

用纯化的血清素/血清胺(ST)抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入血清素/血清胺(ST),再与HRP标记的血消素/血清胺(ST)抗体结合，形成抗体-抗原-的标抗体梵合物，经过彻底洗涤后加底物TMB显色。

TMB在HRP酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。

颜色的深浅和样品中的血清索/血清胺(ST)呈正相关。

用假标仪在4SO AM波长下测定吸光度(OD值)，通过标准曲线计算样品中血清素/血清胺(SD浓度。

试剂盒性能：1.灵敏度：Z山小的检测浓度小于1号标准品。

稀释度的线性。

样品线性回归与预期浓度相关系数尺值为Oqqo2∙特异性：不与其它细胞因子反应。

3.重且性：板内、板间变异系数均小于工O%∙标本要求：1 .标本采集后尽早进行提取，提取按相关文献进行，提取后应尽快进行实验。

若不能马上进行试验，可将标本放于-2。

C 保存，但应避免反完冻融2 .不能检测含NaN3的样品，因NaN3抑制辣根过氧化物陶的(HRP)活性。

样本处理及要求，1■.血清：全血标本请于室温放置2小时或4C过夜后于工Og离心2。

分钟，取上清即可检测，或将标本放于-WC或-8OC保存，但应避免反复冻融。

2∙血浆：可用EDTA或肝素作为抗凝剂，标本采集后30分钟内于2-8。

ClRog离心20分钟，或将标本放于-3七或-8O c C 保存，但应避免反复冻融，3 .组织匀浆：用预冷的PBS9Ql∙M,pH=7.4)冲洗组织，去除残留血液(匀浆中裂解的红细胞会影响测量结果)，称重后将组织剪碎。

将剪碎的组织与对应体积的PBS(一般按1:9的重量体积比，比如Ig的组织样品对应9M L的PBS,具体体积可根据实验需要适当调整，并做好记录。

推荐在PBS中加入蛋白酶抑制剂)加入玻璃匀浆器中，于冰上充分研磨。

为了进一步裂解组织细胞，可以对匀浆液进行超声破碎，或反熨冻融。

酵母表达载体pPICZ手册

pPICZ A, B, and CPichia expression vectors for selection onZeocin™ and purification of recombinant proteins Catalog no. V190-20Rev. Date: 7 July 2010Manual part no. 25-0148MAN00000034User ManualiiTable of ContentsKit Contents and Storage (iv)Accessory Products (v)Introduction (1)Overview (1)Methods (3)Cloning into pPICZ A, B, and C (3)Pichia Transformation (9)Expression in Pichia (13)Purification (15)Appendix (17)Recipes (17)Zeocin™ (19)Map and Features of pPICZ A, B, and C (21)Lithium Chloride Transformation Method (23)Construction of In Vitro Multimers (24)Technical Support (32)Purchaser Notification (33)References (34)iiiKit Contents and StorageContents The following components are included with Catalog no. V190–20. Note that thepPICZ expression vectors are supplied in suspension.Component QuantityCompositionpPICZ A Expression Vector 20 μg 40 μl of 0.5 μg/μl vector in10 mM Tris–HCl, 1 mM EDTA,pH 8.0pPICZ B Expression Vector 20 μg 40 μl of 0.5 μg/μl vector in10 mM Tris–HCl, 1 mM EDTA,pH 8.0pPICZ C Expression Vector 20 μg 40 μl of 0.5 μg/μl vector in10 mM Tris–HCl, 1 mM EDTA,pH 8.0GS115/pPICZ/lacZ Positive1 stab --Control strainShipping/Storage The components included with Catalog no. V190–20 are shipped on wet ice.Upon receipt, store as directed below.For long-term storage of your positive control stab strain, we recommendpreparing a glycerol stock immediately upon receipt and storing at –80°C.Component ShippingStorage pPICZ A Expression Vector Wet ice Store at –20°CpPICZ B Expression Vector Wet ice Store at –20°CpPICZ C Expression Vector Wet ice Store at –20°CGS115/pPICZ/lacZ positive control strain Wet ice Store at 4°CivAccessory ProductsAdditional ProductsThe products listed in this section are intended for use with the pPICZ vectors.For more information, visit our web site at or contactTechnical Support (page 32).Product QuantityCatalogno. X-33 Pichia strain 1 stab C180-00GS115 Pichia strain 1 stab C181-00KM71H Pichia strain 1 stab C182-00SMD1168H Pichia strain 1 stab C184-00pPICZα A, B, and C 20 μg each V195-20pPIC6α A,B, and C 20 μg each V215-20pPIC6 A, B, and C 20 μg each V210-20pPIC6 Starter Kit 1 kit K210-01Original Pichia Expression Kit 1 kit K1710-01EasySelect™Pichia Expression Kit 1 kit K1740-01Pichia EasyComp™ Transformation Kit 1 kit K1730-01Pichia Protocols 1 book G100-01PureLink™ Gel Extraction Kit 50 preps250 prepsK2100–12K2100–25S.N.A.P ™ Gel Purification Kit 25 preps K1999–25PureLink™ Quick Plasmid Miniprep Kit 50 preps250 prepsK2100–10K2100–11PureLink™ HiPure Plasmid Midiprep Kit 25 preps50 prepsK2100–04K2100–13One Shot® TOP10 (chemically competent E. coli) 10 reactions20 reactionsC4040–10C4040–03One Shot® TOP10 Electrocompetent E. Coli 10 reactions20 reactionsC4040-50C4040-52TOP10 Electrocomp™ Kits 20 reactions C664–55Positope™ Control Protein 5 μg R900-50CIAP (Calf Intestinal Alkaline Phosphatase) 1,000 units 18009–019T4 DNA Ligase 100 units500 units15224–01715224–025Zeocin™ 1g5 gR250-01R250-05β-Gal Assay Kit 1 kit K1455-01β-Gal Staining Kit 1 kit K1465-01E-Gel® Agarose Gels E-Gel® Agarose Gels are bufferless, pre-cast agarose gels designed for fast, convenient electrophoresis of DNA samples. E-Gel® agarose gels are available in different agarose percentage and well format for your convenience.For more details on these products, visit our web site at or contact Technical Support (page 32).Continued on next pagevAccessory Products, ContinuedZeocin™Zeocin™ may be obtained from Invitrogen (see above). For your convenience, the drug is prepared in autoclaved, deionized water and available in 1.25 ml aliquotsat a concentration of 100 mg/ml. The stability of Zeocin™ is guaranteed for sixmonths if stored at –20°C.Detection of Fusion Protein A number of antibodies are available from Invitrogen to detect expression ofyour fusion protein from the pPICZ vector. Horseradish peroxidase (HRP)-conjugated antibodies allow one-step detection in Western blots usingcolorimetric or chemiluminescent detection methods. The amount of antibodysupplied is sufficient for 25 Western Blots.Antibody Epitope Catalogno.Anti-myc R950–25 Anti-myc-HRPDetects the 10 amino acid epitopederived from c-myc (Evans et al., 1985):EQKLISEEDLR951–25Anti-His(C-term) R930–25Anti-His(C-term)-HRPDetects the C-terminal polyhistidine(6xHis) tag (requires the free carboxylgroup for detection) (Lindner et al., 1997):HHHHHH-COOHR931–25Purification of Fusion Protein The polyhistidine (6xHis) tag allows purification of the recombinant fusionprotein using metal-chelating resins such as ProBond™. Ordering information forProBond™ resin is provided below.Product QuantityCatalogno. ProBond™ Purification System 6 purifications K850–01ProBond ™ Purification System with Anti-myc-HRP Antibody1 Kit K852–01ProBond ™ Purification System with Anti-His(C-term)-HRP Antibody1 Kit K853–01ProBond™ Nickel-Chelating Resin 50 ml150 mlR801–01R801–15Purification Columns 50 each R640–50viIntroductionOverviewIntroduction pPICZ A, B, and C are 3.3 kb expression vectors used to express recombinantproteins in Pichia pastoris. Recombinant proteins are expressed as fusions to aC-terminal peptide containing the c-myc epitope and a polyhistidine (6xHis) tag.The vector allows high-level, methanol inducible expression of the gene ofinterest in Pichia, and can be used in any Pichia strain including X33, GS115,SMD1168H, and KM71H. pPICZ contains the following elements:•5′ fragment containing the AOX1 promoter for tightly regulated, methanol-induced expression of the gene of interest (Ellis et al., 1985; Koutz et al., 1989;Tschopp et al., 1987a)•Zeocin™ resistance gene for selection in both E. coli and Pichia (Baron et al.,1992; Drocourt et al., 1990)•C-terminal peptide containing the c-myc epitope and a polyhistidine (6xHis)tag for detection and purification of a recombinant fusion protein (if desired)•Three reading frames to facilitate in-frame cloning with the C-terminalpeptideReference Sources The pPICZ A, B, and C expression vectors may be used with the Original Pichia Expression Kit, and are included in the EasySelect™Pichia Expression Kit (see page v for ordering information). Additional general information about recombinant protein expression in Pichia pastoris is provided in the manuals for the Original Pichia Expression Kit and the EasySelect™Pichia Expression Kit. For more information about the Original Pichia Expression Kit, the EasySelect™Pichia Expression Kit, or their manuals, visit our web site at or contact Technical Support (page 32).More detailed information and protocols dealing with Pichia pastoris may also be found in the following general reference:Higgins, D. R., and Cregg, J. M. (1998) Pichia Protocols. In Methods in Molecular Biology, Vol. 103. (J. M. Walker, ed. Humana Press, Totowa, NJ) (see page v for ordering information).Recommended Pichia Host Strain We recommend using the X-33 Pichia strain as the host for expression of recombinant proteins from pPICZ. Other Pichia strains including GS115, KM71H, and SMD1168H are suitable. The X-33 Pichia strain and other strains are available from Invitrogen (see page v for ordering information). The X-33 Pichia strain has the following genotype and phenotype:Genotype: Wild-typePhenotype: Mut+1Overview, ContinuedExperimental Overview The following table describes the basic steps needed to clone and express your gene of interest in pPICZ.Step Action1 Propagate pPICZ A, B, and C by transformation into a rec A, end A1E. coli strain such as TOP10, DH5 , or JM109.2 Develop a cloning strategy and ligate your gene into one of the pPICZvectors in frame with the C-terminal tag.3 TransformintoE. coli and select transformants on Low Salt LB platescontaining 25 μg/ml Zeocin™.4 Analyze 10–20 transformants by restriction mapping or sequencing toconfirm in-frame fusion of your gene with the C-terminal tag.5 Purify and linearize the recombinant plasmid for transformation intoPichia pastoris.6 TransformyourPichia strain and plate onto YPDS plates containing the appropriate concentration of Zeocin™.7 Select for Zeocin™-resistant transformants.8 Optimize expression of your gene.9 Purify your fusion protein on metal-chelating resin (i.e. ProBond™).Continued on next page2MethodsCloning into pPICZ A, B, and CIntroduction The pPICZ vector is supplied with the multiple cloning site in three readingframes (A, B, and C) to facilitate cloning your gene of interest in frame with theC-terminal peptide containing the c-myc epitope and a polyhistidine (6xHis) tag.Use the diagrams provided on pages 5–7 to help you design a strategy to cloneyour gene of interest in frame with the C-terminal peptide. Generalconsiderations for cloning and transformation are discussed in this section.General Molecular Biology Techniques For assistance with E. coli transformations, restriction enzyme analysis, DNA biochemistry, and plasmid preparation, refer to Molecular Cloning: A Laboratory Manual (Sambrook et al., 1989) or Current Protocols in Molecular Biology (Ausubel et al., 1994).E. coli Strain Many E. coli strains are suitable for the propagation of the pPICZ vectorsincluding TOP10, JM109, and DH5 . We recommend that you propagate thepPICZ vectors in E. coli strains that are recombination deficient (rec A) andendonuclease A deficient (end A).For your convenience, TOP10 E. coli are available as chemically competent orelectrocompetent cells from Invitrogen (page v).Transformation Method You may use any method of choice for transformation. Chemical transformation is the most convenient for many researchers. Electroporation is the most efficient and the method of choice for large plasmids.Maintenance of Plasmids The pPICZ vectors contain the Zeocin™ resistance (Sh ble) gene to allow selection of the plasmid using Zeocin™. To propagate and maintain the pPICZ plasmids, we recommend using the following procedure:e 10 ng of your vector to transform a rec A, end A E. coli strain like TOP10,DH5 , JM109, or equivalent (see above).2.Select transformants on Low Salt LB plates containing 25 μg/ml Zeocin™ (seepage 17 for a recipe).3.Prepare a glycerol stock from each transformant containing plasmid forlong-term storage (see page 8).Continued on next page3Cloning into pPICZ A, B, and C, ContinuedGeneral Considerations The following are some general points to consider when using pPICZ to express your gene of interest in Pichia:•The codon usage in Pichia is believed to be similar to Saccharomyces cerevisiae.•Many Saccharomyces genes have proven to be functional in Pichia.•The premature termination of transcripts because of "AT rich regions" has been observed in Pichia and other eukaryotic systems (Henikoff & Cohen, 1984; Irniger et al., 1991; Scorer et al., 1993; Zaret & Sherman, 1984). If you have problems expressing your gene, check for premature termination by northern analysis and check your sequence for AT rich regions. It may be necessary to change the sequence in order to express your gene (Scorer et al., 1993).•The native 5´ end of the AOX1 mRNA is noted in the diagram for each multiple cloning site. This information is needed to calculate the size of the expressed mRNA of the gene of interest if you need to analyze mRNA for any reason.Cloning Considerations For proper initiation of translation, your insert should contain an initiation ATG codon as part of a yeast consensus sequence (Romanos et al., 1992). An example of a yeast consensus sequence is provided below. The ATG initiation codon is shown underlined.(G/A)NNATG GTo express your gene as a recombinant fusion protein, you must clone your gene in frame with the C-terminal peptide containing the c-myc epitope and the polyhistidine tag. The vector is supplied in three reading frames to facilitate cloning. Refer to the diagrams on pages 5–7 to develop a cloning strategy.If you wish to express your protein without the C-terminal peptide, be sure to include a stop codon.Construction of Multimeric Plasmids pPICZ A, B, and C contain unique Bgl II and Bam H I sites to allow construction of plasmids containing multiple copies of your gene. For information on how to construct multimers, refer to pages 24–31.Continued on next page4Multiple CloningSite of pPICZ A Below is the multiple cloning site for pPICZ A. Restriction sites are labeled to indicate the cleavage site. The boxed nucleotides indicate the variable region.The multiple cloning site has been confirmed by sequencing and functionaltesting.You can download the complete sequence of pPICZ A from our web site at or by contacting Technical Support (see page 32).For a map and a description of the features of pPICZ, refer to the Appendix(pages 21–22). AAT AGC GCC GTC GAC CAT CAT CAT CAT CAT CAT TGTTCCTCAG TTCAAGTTGG GCACTTACGA GAAGACCGGT CTTGCTAGAT TCTAATCAAG AGGATGTCAG AATGCCATTT GCCTGAGAGA TGCAGGCTTC ATTTTTGATA CTTTTTTATTTGTAACCTAT ATAGTATAGG ATTTTTTTTG TCATTTTGTT 1218Asn Ser Ala Val Asp His His His His His His ***3´ AOX1 priming site TGA GTTTTAGCCT TAGACATGAC AACCTTTTTT TTTATCATCA TTATTAGCTT ACTTTCATAA TTGCGACTGG TTCCAATTGA CAAGCTTTTG ATTTTAACGA CTTTTAACGA CAACTTGAGA AGATCAAAAA ACAACTAATT ATTCGAAACG AGGAATTCAC GTGGCCCAGC CGGCCGTCTC GGATCGGTAC CTCGAGCCGC GGCGGCCGCC AGCTT GGGCCC GAA CAA AAA CTC ATC TCA GAA GAG GAT CTG 811Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 5´ AOX1 priming sitemyc epitope3´polyadenylation sitePolyhistidine tag5´ end of AOX1 mRNA Sfu I Eco R I Pml I Sfi I Bsm B I Asp 718 I Kpn I Xho ISac II Not I Apa I 104210981158871931991Continued on next pageMultiple CloningSite of pPICZ B Below is the multiple cloning site for pPICZ B. Restriction sites are labeled to indicate the cleavage site. The boxed nucleotides indicate the variable region.The multiple cloning site has been confirmed by sequencing and functionaltesting.You can download the complete sequence of pPICZ B from our web site at or by contacting Technical Support (see page 32).For a map and a description of the features of pPICZ, refer to the Appendix(pages 21–22). AAT AGC GCC GTC GAC CAT CAT CAT CAT CAT CAT TGA GTTTGTAGCC TTAGACATGA CTGTTCCTCA GTTCAAGTTG GGCACTTACG AGAAGACCGG TCTTGCTAGA TTCTAATCAA GAGGATGTCA GAATGCCATT TGCCTGAGAG ATGCAGGCTT CATTTTTGAT ACTTTTTTAT TTGTAACCTA TATAGTATAG GATTTTTTTT GTCATTTTGT TTC 1216Asn Ser Ala Val Asp His His His His His His ***3´ AOX1 priming site TGA GTTTGTAGCC TTAGACATGA AACCTTTTTT TTTATCATCA TTATTAGCTT ACTTTCATAA TTGCGACTGG TTCCAATTGA CAAGCTTTTG ATTTTAACGA CTTTTAACGA CAACTTGAGA AGATCAAAAA ACAACTAATTATTCGAAACG AGGAATTCAC GTGGCCCAGC CGGCCGTCTC GGATCGGTAC CTCGAGCCGC GGCGGCCGCC AGCTT TCTA GAA CAA AAA CTC ATC TCA GAA GAG GAT CTG 811Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 5´ AOX1 priming sitemyc epitope3´ polyadenylation site Polyhistidine tag5´ end of AOX1 mRNA Sfu I Eco R I Pml I Sfi I Bsm B I Asp 718 I Kpn I Xho ISac II Not I Xba I 104010961156871931991Continued on next pageMultiple CloningSite of pPICZ C Below is the multiple cloning site for pPICZ C. Restriction sites are labeled to indicate the cleavage site. The boxed nucleotides indicate the variable region.The multiple cloning site has been confirmed by sequencing and functionaltesting.You can download the complete sequence of pPICZ C from our web site at or by contacting Technical Support (see page 32).For a map and a description of the features of pPICZ, refer to the Appendix(pages 21–22). AAT AGC GCC GTC GAC CAT CAT CAT CAT CAT CAT TGA GTTTGTAGCC TTAGACATGA CTGTTCCTCA GTTCAAGTTG GGCACTTACG AGAAGACCGG TCTTGCTAGA TTCTAATCAA GAGGATGTCA GAATGCCATT TGCCTGAGAG ATGCAGGCTT CATTTTTGAT ACTTTTTTAT TTGTAACCTA TATAGTATAG GATTTTTTTT GTCATTTTGT TTC 1217Asn Ser Ala Val Asp His His His His His His ***3´ AOX1 priming siteTGA GTTTGTAGCC TTAGACATGA AACCTTTTTT TTTATCATCA TTATTAGCTT ACTTTCATAA TTGCGACTGG TTCCAATTGA CAAGCTTTTG ATTTTAACGA CTTTTAACGA CAACTTGAGA AGATCAAAAA ACAACTAATT ATTCGAAACG AGGAATTCAC GTGGCCCAGC CGGCCGTCTC GGATCGGTAC CTCGAGCCGC GGCGGCCGCC AGCTT ACGTA GAA CAA AAA CTC ATC TCA GAA GAG GAT CTG 811Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 5´ AOX1 priming sitemyc epitope3´ polyadenylation site Polyhistidine tag5´ end of AOX1 mRNA Sfu I Eco R I Pml I Sfi I Bsm B I Asp 718 I Kpn I Xho I Sac II Not I Sna B I 104110971157871931991Continued on next pageE. coli Transformation Transform your ligation mixtures into a competent rec A, end A E. coli strain(e.g. TOP10, DH5, JM109) and select on Low Salt LB agar plates containing25 μg/ml Zeocin™ (see below). Note that there is no blue/white screening for the presence of insert with pPICZ A, B, or C. Once you have obtained Zeocin™-resistant colonies, pick 10 transformants and screen for the presence and orientation of your insert.Important To facilitate selection of Zeocin™-resistant E. coli, the salt concentration of the medium must remain low (<90 mM) and the pH must be 7.5. Prepare Low Salt LB broth and plates using the recipe in the Appendix, page 17.Failure to lower the salt content of your LB medium will result in non-selection due to inhibition of the drug.C We recommend that you sequence your construct to confirm that your gene is in the correct orientation for expression and cloned in frame with the C-terminal peptide (if desired). Refer to the diagrams on pages 5–7 for the sequences and location of the priming sites.Preparing a Glycerol Stock Once you have identified the correct clone, be sure to purify the colony and make a glycerol stock for long-term storage. It is also a good idea to keep a DNA stock of your plasmid at –20°C.1.Streak the original colony out on an Low Salt LB plate containing 25 μg/mlZeocin™. Incubate the plate at 37°C overnight.2.Isolate a single colony and inoculate into 1–2 ml of Low Salt LB containing25 μg/ml Zeocin™.3.Grow the culture to mid-log phase (OD600 = 0.5–0.7).4.Mix 0.85 ml of culture with 0.15 ml of sterile glycerol and transfer to acryovial.5.Store at –80°C.Plasmid Preparation Once you have cloned and sequenced your insert, generate enough plasmid DNA to transform Pichia (5–10 μg of each plasmid per transformation). We recommend isolating plasmid DNA using the PureLink™ Quick Plasmid Miniprep Kit or the PureLink™ HiPure Plasmid Midiprep Kit (page v), or CsCl gradient centrifugation.Once you have purified plasmid DNA, proceed to Pichia Transformation, next page.Pichia TransformationIntroduction You should now have your gene cloned into one of the pPICZ vectors. Yourconstruct should be correctly fused to the C-terminal peptide (if desired). Thissection provides general guidelines to prepare plasmid DNA, transform yourPichia strain, and select for Zeocin™-resistant clones.Zeocin™ Selection We generally use 100 μg/ml Zeocin™ to select for transformants when using the X-33 Pichia strain. If you are transforming your pPICZ construct into anotherPichia strain, note that selection conditions may vary. We recommendperforming a dose response curve to determine the appropriate concentration ofZeocin™ to use for selection of transformants in your strain.Method of Transformation We do not recommend spheroplasting for transformation of Pichia with plasmids containing the Zeocin™ resistance marker. Spheroplasting involves removal of the cell wall to allow DNA to enter the cell. Cells must first regenerate the cell wall before they are able to express the Zeocin™ resistance gene. For this reason, plating spheroplasts directly onto selective medium containing Zeocin™ does not yield any transformants.We recommend electroporation for transformation of Pichia with pPICZ A, B, or C. Electroporation yields 103 to 104 transformants per μg of linearized DNA and does not destroy the cell wall of Pichia. If you do not have access to an electroporation device, use the LiCl protocol on page 23 or the Pichia EasyComp™Transformation Kit available from Invitrogen (see below).PichiaEasyComp™Transformation Kit If you wish to perform chemical transformation of your Pichia strain with pPICZ A, B, or C, the Pichia EasyComp™ Transformation Kit is available from Invitrogen (see page v for ordering information). The Pichia EasyComp™ Transformation Kit provides reagents to prepare 6 preparations of competent cells. Each preparation will yield enough competent cells for 20 transformations. Competent cells may be used immediately or frozen and stored for future use. For more information, visit our web site at or contact Technical Support (page 32).Important Since pPICZ does not contain the HIS4 gene, integration can only occur at the AOX1 locus. Vector linearized within the 5´ AOX1 region will integrate by gene insertion into the host 5´ AOX1 region. Therefore, the Pichia host that you use will determine whether the recombinant strain is able to metabolize methanol (Mut+) or not (Mut S). To generate a Mut+ recombinant strain, you must use a Pichia host that contains the native AOX1 gene (e.g. X-33, GS115, SMD1168H). If you wish to generate a Mut S recombinant strain, then use a Pichia host that has a disrupted AOX1 gene (i.e. KM71H).Continued on next pageHis4 Host Strains Host strains containing the his4 allele (e.g. GS115) and transformed with thepPICZ vectors require histidine when grown in minimal media. Add histidine toa final concentration of 0.004% to ensure growth of your transformants.The pPICZ vectors do not contain a yeast origin of replication. Transformantscan only be isolated if recombination occurs between the plasmid and the Pichiagenome.Materials Needed You will need the following items:Note: Inclusion of sorbitol in YPD plates stabilizes electroporated cells as they appear tobe somewhat osmotically sensitive.•5–10 μg pure pPICZ containing your insert•YPD Medium•50 ml conical polypropylene tubes• 1 liter cold (4°C) sterile water (place on ice the day of the experiment)•25 ml cold (4°C) sterile 1 M sorbitol (place on ice the day of the experiment)•30°C incubator•Electroporation device and 0.2 cm cuvettes•YPDS plates containing the appropriate concentration of Zeocin™ (seepage 18 for recipe)Linearizing YourpPICZ ConstructTo promote integration, we recommend that you linearize your pPICZ constructwithin the 5′ AOX1 region. The table below lists unique sites that may be used tolinearize pPICZ prior to transformation. Other restriction sites are possible.Note that for the enzymes listed below, the cleavage site is the same for versionsA, B, and C of pPICZ. Be sure that your insert does not contain the restriction siteyou wish to use to linearize your vector.Enzyme Restriction Site (bp) SupplierSac I 209 ManyPme I 414 New England BiolabsBst X I 707 ManyRestriction Digest 1.Digest ~5–10 μg of plasmid DNA with one of the enzymes listed above.2.Check a small aliquot of your digest by agarose gel electrophoresis forcomplete linearization.3.If the vector is completely linearized, heat inactivate or add EDTA to stopthe reaction, phenol/chloroform extract once, and ethanol precipitate using1/10 volume 3 M sodium acetate and 2.5 volumes of 100% ethanol.4.Centrifuge the solution to pellet the DNA, wash the pellet with 80% ethanol,air-dry, and resuspend in 10 μl sterile, deionized water. Use immediately orstore at –20°C.Continued on next pagePreparation of Pichia for Electroporation Follow the procedure below to prepare your Pichia pastoris strain for electroporation.1. Grow 5 ml of your Pichia pastoris strain in YPD in a 50 ml conical tube at30°C overnight.2. Inoculate 500 ml of fresh medium in a 2 liter flask with 0.1–0.5 ml of theovernight culture. Grow overnight again to an OD600 = 1.3–1.5.3. Centrifuge the cells at 1500 × g for 5 minutes at 4°C. Resuspend the pelletwith 500 ml of ice-cold (0–4°C), sterile water.4. Centrifuge the cells as in Step 3, then resuspend the pellet with 250 ml ofice-cold (0–4°C), sterile water.5. Centrifuge the cells as in Step 3, then resuspend the pellet in 20 ml of ice-cold (0–4°C) 1 M sorbitol.6. Centrifuge the cells as in Step 3, then resuspend the pellet in 1 ml of ice-cold(0–4°C) 1 M sorbitol for a final volume of approximately 1.5 ml. Keep the cells on ice and use that day. Do not store cells.Transformation by Electroporation 1.Mix 80 μl of the cells from Step 6 (above) with 5–10 μg of linearized pPICZDNA (in 5–10 μl sterile water) and transfer them to an ice-cold (0–4°C)0.2 cm electroporation cuvette.2.Incubate the cuvette with the cells on ice for 5 minutes.3.Pulse the cells according to the parameters for yeast (Saccharomycescerevisiae) as suggested by the manufacturer of the specific electroporation device being used.4.Immediately add 1 ml of ice-cold 1 M sorbitol to the cuvette. Transfer thecuvette contents to a sterile 15 ml tube.5.Let the tube incubate at 30°C without shaking for 1 to 2 hours.6.Spread 50-200 μl each on separate, labeled YPDS plates containing theappropriate concentration of Zeocin™.7.Incubate plates for 2–3 days at 30°C until colonies form.8.Pick 10–20 colonies and purify (streak for single colonies) on fresh YPD orYPDS plates containing the appropriate concentration of Zeocin™.Continued on next pageGenerally, several hundred Zeocin™-resistant colonies are generated using theprotocol on the previous page. If more colonies are needed, the protocol may bemodified as described below. Note that you will need ~20, 150 mm plates withYPDS agar containing the appropriate concentration of Zeocin™.1. Set up two transformations per construct and follow Steps 1 through 5 ofthe Transformation by Electroporation protocol, page 11.2. After 1 hour in 1 M sorbitol at 30°C (Step 5, previous page), add 1 ml YPDmedium to each tube.3. Shake (~200 rpm) the cultures at 30°C.4. After 1 hour, take one of the tubes and plate out all of the cells by spreading200 μl on 150 mm plates containing the appropriate concentration ofZeocin™.5. Optional: Continue incubating the other culture for three more hours (for atotal of four hours) and then plate out all of the cells by spreading 200 μl on150 mm plates containing the appropriate concentration of Zeocin™.6. Incubate plates for 2–4 days at 30°C until colonies form.Mut Phenotype If you used a Pichia strain containing a native AOX1 gene (e.g. X-33, GS115,SMD1168H) as the host for your pPICZ construct, your Zeocin™-resistanttransformants will be Mut+. If you used a strain containing a deletion in theAOX1 gene (e.g. KM71H), your transformants will be Mut S.If you wish to verify the Mut phenotype of your Zeocin™-resistant transformants,you may refer to the general guidelines provided in the EasySelect™PichiaExpression Kit manual or the Original Pichia Expression Kit manual or topublished reference sources (Higgins & Cregg, 1998).You are now ready to test your transformants for expression of your gene ofinterest. See Expression in Pichia, next page.。

毕赤酵母表达系统使用心得

毕⾚酵母表达系统使⽤⼼得Pichia酵母表达系统使⽤⼼得甲醇酵母表达系统有不少优点，其中以Invitrogen公司的Pichia酵母表达系统最为⼈熟知，并⼴泛应⽤于外源蛋⽩的表达。

虽然说酵母表达操作简单表达量⾼，但是在实际操作中，并不是每个外源基因都能顺利得到⾼表达的。

不少⼈在操作中会遇到这样那样的问题，收集了部分⽤户在使⽤EasySelect Pichia Expression System这个被誉为最简单的毕⾚酵母表达的经典试剂盒过程中的⼼得体会。

其中Xiang Yang是来⾃美国乔治城⼤学（Georgetown University）Lombardi癌症中⼼（Lombardi Cancer Center），部分⽤户来⾃国内。

+ 表⽰优胜于；- 表⽰不如；= 表⽰差不多EasySelect Pichia Expression System产品性能：优点——使⽤简单，表达量⾼，His-tag便于纯化缺点——酵母表达蛋⽩有时会出现蛋⽩切割问题全⾯产品报告及⼼得体会：巴斯德毕⾚酵母（Pichia pastoris ）是⼀种能⾼效表达重组蛋⽩的酵母品种，⼀⽅⾯由于其是属于真核⽣物，因此表达出来的蛋⽩可以进⾏糖基化修饰，另⼀⽅⾯毕⾚酵母⽣长速度快，可以将表达的蛋⽩分泌到培养基中，⽅便蛋⽩纯化。

毕⾚酵母表达载体pPICZ 在多克隆位点（MCR ）3'端带有his-tag 和c-myc epitopes ，这些tag 有利于常规检测和纯化，⽽且在MCR5'端引⼊了alpha factor （α-factor ）⽤以增加表达，并且在表达后α-factor 可以⾃动被切除。

在进⾏克隆的时候，如果你选择的是EcoRI ，那么只需在⽬标蛋⽩中增加两个氨基酸序列即可完成。

另外pPICZ 系列选⽤的是Zeocin 抗⽣素作为筛选标记，⽽诱导表达的载体需要甲醇——甲醇⽐⼀般⽤于⼤肠杆菌表达诱导使⽤的IPTG 便宜。

所有质粒载体汇总

pEZZ18 pkk232-8,pkk 233-3,pACYC184,pBR322,pUC119 pTYB1,pTYB2,pTYB4,pTYB11 pBlueScript SK（+）,pBlueScript SK（-） pLLP ompA, pINIIIompA, pMBP-P ,pMBP-C, 大肠杆菌冷激质粒： pColdI pColdII pColdIII pColdTF 原核共表达质粒：pACYCduet1,pETduet-1,pCDFduet-1，pRSFduet-1 Takara公司大肠杆菌分子伴侣： pG-KJE8 pGro7 pKJE7 pGTf2 pTf16 大肠杆菌宿主细胞： DH5a JM101 JM103 JM105 JM107 JM109 JM110 Top10 Top10F BL21（DE3） HB101 ER2529 E2566 C2566 MG1655 XL-10gold XL blue M15 JF1125 K802 SG1117 BL21（AI） BL21（DE3）plysS TG1 TB1 DH5a（pir） Tuner（DE3） Bl21 codonplusRIPL Novablue（DE3） Rosetta Rosetta（DE3） Rosetta（DE3）plys Rosetta-gami（DE3） RosettagamiB（DE3）, Rosetta-gamiB（DE3）plysS Orgami（DE3） OrgamiB（DE3） HMS174（DE3）植物表达/RNAi载体农杆菌pBI121,pBI121-GFP,pBI101,pBI221,pSN1301, pUN1301,pRTL2 , pRTL2-GFP , pRTL2-CFP, pRTL2-RFP , pRTL2YFP,pCAMBIA 1300, 1301, 1302,1303,1304,1305, 1381Z,1391Z,2300, 2301,3300,3301,pCAMBIA super1300,pCAMBIA super1300GFP,pPZP212,pPZP2121,pPZP212-GFP,pGDG,RNAi载体pART27,pHANNIBAL,pKANNIBAL, pFGC5941,pTCK303, pTRV1,pTRV2, T-DNA插入载体（随机突变体库）pSKI015,pSKI074,真菌ATMT载体pBIG2RHPH2-GUS-GFP,pBHt1 枯草芽孢杆菌表达载体pWB980,pHT43,pHP13,pHP43, pBE2,pMUTIN4,pUB110,pE194,pMA5, pMK3,pMK4,pHT304,pHY300PLK, pBest502,pDG1363,pSG1154,pAX01, pSAS144,pDL,pDG148-stu,pDG641, pAL12,pUCX05-bgaB,pHT01，配套菌株BS 168,WB600,WB800,WB700, WB800N,1012,FZB42,1A747,广宿主质粒pVLT33

酵母表达系统步骤

酵母表达系统步骤毕赤酵母表达系统步骤（参考Invitrogen公司说明书）：一、pPICZαA、B、C质粒以及DH5α菌株的保存1取0.5μl pPICZα A、B、C质粒，热击转化DH5α，在低盐LB （含有25μg/ml Zeocin）的平板上37℃培养过夜。

2挑取转化子，甘油保存。

二、载体构建1将目的基因构建到pPICZα载体上，转化DH5α，用Zeocin筛选转化子。

2提质粒酶切鉴定或PCR鉴定3载体测序测序可用α-Factor引物或5’AOX1引物，3’AOX1引物三、线性化DNA1提取足够量的质粒DNA（一次转化至少需要5-10μg质粒）2 酶切线性化10μg构建好的载体，同时酶切空载体做对照，根据载体选择线性化酶切位点（样品分管酶切），pPICZα载体在5’AOX1区域有三个酶切位点可选择：SacI、PmeI、BstXI3 取1-2μl酶切产物跑电泳，确定是否酶切完全；4 过柱纯化线性化质粒（用50μl EB洗脱）；四、线性化DNA的去磷酸化处理线性化质粒43μlCIAP Buffer 5μlCIAP酶2μl四、总体积为50μl的样品37℃ 1h，过柱纯化，用30μl ddH2O 洗脱；五、感受态细胞的制备实验前准备：无抗性YPD平板一个、无抗生素液体YPD培养基，100μg/ml Zeocin YPD 平板和液体、50ml离心管两个、500ml预冷的无菌水、20ml 1M 山梨醇（灭菌预冷的），0.2cm预冷的电击杯；1YPD平板划线培养菌，30℃培养2-3d；250ml三角瓶中，加入5ml YPD，挑取酵母单菌落，30℃培养过夜；3吸取0.5ml菌液，加入至含有200ml新鲜YPD的1L三角瓶中，30℃，225rpm/min培养至OD值1.3-1.5；41500g，4℃离心5min收集菌体；540ml冰预冷的无菌水重悬沉淀；61500g，4℃,5min；730ml无菌水重悬；81500g，4℃,5min；910ml 1M 山梨醇重悬；101500g，4℃,5min；11加入1ml山梨醇，重悬冰上放置，直接做转化，或加入灭菌甘油每管80ul分装，冻存于-80℃(长时间保存会影响转化效率)；六、电击转化15-10μg线性化DNA（20μl<）与80ul上述感受态细胞混合，转移至预冷的0.2cm电击杯中（点击条件：电压1.5kV；电容25μF；电阻200Ω，电击时间为4～10msec）；2冰上放置5min3电击（按生产厂商提供的适合酵母用的参数）4迅速加入1ml预冷的1M 山梨醇，转移至1.5ml EP管中530℃静置培养1-2h（如果要增加存活率，获得更多的转化克隆，可在30℃静置培养1h后，加入1mlYPD培养基，30℃200rpm培养1h后取部分涂布与不同浓度抗生素的平板）6取50、100、200ul分别涂布于含有Zeocin的YPD平板，30℃培养2-10 d至有菌落出现；7如果要筛选多拷贝转化子，将转化克隆混合在一起，涂布在Zeocin 浓度为500、1000、2000μg/ml的YPD平板，培养2-3d。

pPICZ A酵母表达载体说明

pPICZ A编号载体名称北京华越洋生物VECT2440 pPICZ ApPICZA载体基本信息出品公司: Invitrogen载体名称: pPICZA, p PICZ A质粒类型: 毕赤酵母蛋白表达载体表达水平: 高拷贝启动子: AOX1克隆方法: 多克隆位点，限制性内切酶载体大小: 3329 b p5' 测序引物及序列: 5´ A OX1:5´-‐GACTGGTTCCAATTGACAAGC-‐3´ 3' 测序引物及序列: 3´ A OX1:5´-‐GCAAATGGCATTCTGACATCC-‐3´ 载体标签: C-‐Myc, C-‐His载体抗性: Zeocin 博来霉素筛选标记: HIS4备注: 插入基因是必需包含起始密码子ATG。

产品目录号: -‐-‐稳定性: 稳定 Stable组成型: 组成型 Constitutive病毒/非病毒: 非病毒pPICZA载体质粒图谱和多克隆位点信息pPICZA多克隆位点pPICZA载体简介pPICZ A，B和C是3.3 k b的毕赤酵母蛋白载体。

表达的重组蛋白是融合蛋白，含有一个C-‐端多肽，多太重含c-‐myc和C-‐His标签。

载体能够在毕赤酵母中利用甲醇诱导的高水平的表达目的蛋白，并且可以用在任何毕赤酵母中，包括X33，GS115菌株，SMD1168H，KM71H。

pPICZ系列载体包含以下元素：•5'片段含有AOX1启动子的严格调控，利用甲醇诱导表达任何感兴趣的基因（Ellis等，1985; Koutz等人，1989;tschopp等人，1987A）。

•Zeocin抗性基因在大肠杆菌和毕赤酵母都能用于筛选（Baron等人,1992; D rocourt等人，1990）。

常用载体及相关通用引物

常用载体及相关通用引物默认分类2008-10-06 13:16:31 阅读512 评论0 字号：大中小载体一端另一端pBlueScriptSK(+) M13F(-77)/M13F(-47)/M13F/T7 M13R(-88)/M13R(-48)/M13R/T3 pcDNA3 H1.3F/pCMV-F /pEGFP-N-5/T7 BGHpcDNA3.1(+) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.0(+)/myc-His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 BGHpcDNA3.1(+)/myc-His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1(+)/myc-His/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/His/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Hygro(-) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Hygro(+) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Hygro/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His-TOPO H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His-TOPO/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Zeo(-) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Zeo(+) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Zeo/CAT H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpColdI DNA p-GEX3’pColdII DNA p-GEX3’pColdIII DNA p-GEX3’pColdIV DNA p-GEX3’pDream2.1 H1.3F/pCMV-F /pEGFP-N-5/T7 SP6pEGFP-N1,2,3 H1.3F/pCMV-F /pEGFP-N-5 pEGFP-N-3’/pEGFP-C-5’/pEGFP-C-3' pET15b T7 T7terpET20b T7 T7terpET22b T7 T7terpET28a T7 T7terpET28b T7 T7terpET30b T7 T7terpFastBac Dual pEGFP-C-3'pFastBac HT A,B,C pEGFP-C-3'pFastBac1 pEGFP-C-3'pGEM-T Easy M13F(-77)/M13F(-47)/M13F/T7 M13R(-88)/M13R(-48)/M13RpGEX-4T-1/3 p-GEX5’ p-GEX3’pGS-21a p-GEX5’ T7terpLenti6/V5-D-TOPO M13F(-47)/M13F/T7 M13R(-48)/M13R/T3pLenti6/V5-GW/lacZ M13F(-47)/M13F/T7 M13R(-48)/M13R/T3pMAL-p2x M13F(-47)/M13F/PBV220R M13R(-48)pMD18-T M13F(-77)/M13F(-47)/M13F M13R(-88)/M13R(-48)/M13RpMD19-T M13F(-77)/M13F(-47)/M13F M13R(-88)/M13R(-48)/M13RpMTBiPV5-His GFP p-GEX3’/ M13F(-47)/M13F M13R(-88)/M13R(-48)/M13R /pEGFP-C-3'/BGH pPCR script M13F(-77)/M13F(-47)/M13F/T7 M13R(-88)/M13R(-48)/M13R/T3pPIC3.5 5’AOX 3’AOXpPIC9 5’AOX 3’AOXpPICZ C/A/B 5’AOX 3’AOXpPICZalpha C/A/B 5’AOX/α-Factor 3’AOXpSecTag2 A,B,C H1.3F/pCMV-F /pEGFP-N-5/T7 BGHpTWIN T7 T7terpUC57 M13F(-77)/M13F(-47)/M13F M13R(-88)/M13R(-48)/M13RpUC18 M13F(-47)/M13F M13R(-48)/M13RpUC19 M13F(-47)/M13F M13R(-48)/M13R其他…… ……注：1）排列顺序为：有左到右==（离插入为点）由远到近；2）若有多个引物，红色的为首选引物：3）选择测序通用引物时，以距离插入位点约50碱基为佳；4）同时有T7，T7ter的载体，单反应首选T7ter；5）同时有T7，pCDNA3.1R的载体，单反应首选pCDNA3.1R ；6）同时有T7，BGH的载体，单反应首选BGH；7）同时有pCMV-F，BGH的载体，单反应首选BGH；。

所有质粒载体汇总

酿酒酵母表达载体pYES2,pYES2/NT,pYES2/CT,pYES3,pYES6, pYCplac22-GFP,酵母载体pAUR123,pRS303TEF,pRS304, pRS305,pRS306,pY13TEF,pY14TEF，pY15TEF，pY16TEF,酵母基因重组表达载体pUG6, pSH47,酵母单杂载体pHISi,pLacZi,pHIS2, pGAD424, 酵母双杂交系统:酿酒酵母Y187, 酿酒酵母AH109；质粒pGADT7,pGBKT7；对照质粒pGBKT7-53，pGBKT7-lam，pGADT7-T，PCL1，酿酒酵母菌株INVSc1,YM4271, AH109,Y187,Y190,毕赤酵母表达载体pPIC9K,pPIC9K-His,pPIC3.5K,pPICZalphaA,B,C,pPICZA,B,C,pGAPZαA,pAO815,pPIC9k-His,pHIL-S1,pPink hc，配套毕赤酵母Pichiapink,毕赤酵母宿主X33，KM71，KM71H，GS115，原核表达载体pQE30,31,32,40,60,61,62,等原核表达载体,包括pET系列，pET-GST，pGEX系列（含GST标签），pMAL系列pMAL-c2x,-c4x,-c4e,-c5x,-p5x,pBAD,pBADHis,pBADmycHis系列，pQE系列,pTrc99a,pTrcHis系列，pBV220,221,222,pTXB系列，pLLP-ompA,pIN-III-ompA （分泌型表达系列）,pQBI63（原核表达带荧光）pET3a, pET 3d, pET 11a, pET 12a, pET 14b, pET 15b, pET 16b, pET 17b, pET 19b, pET 20b, pET 21a,b,d, pET 22b, pET 23a, pET 23b, pET 24a,b, pET 25b, pET 26b, pET 27b, pET 28a,b, pET 29a, pET 30a, pET 31b, pET 32a, pET 35b, pET 38b, pET 39b, pET 40b, pET 41a,b pET 42a, pET 43.1a,b pET 44a, pET 49b pET302,303 pET His,pET Dsb,pET GST,pET Trx pQE2, pQE9 pQE30,31,32, pQE 40 pQE70 pQE80L pQETirs system pRSET-A pRSET-B pRSET-C pGEX4T-1,-2,-3,5x-1,6p-1,6p-2,2tk,3c pBV220,221,222 pTrcHisA,B,C pBAD24,34,43 pBAD HisA,B,C pPinPoint-Xa1,Xa2,Xa3 pMALc2x, p2x pBV220 pGEM Ex1, pGEM7ZF（+）, pTrc99A, pTwin1, pEZZ18 pkk232-8,pkk233-3,pACYC184,pBR322,pUC119 pTYB1,pTYB2,pTYB4,pTYB11 pBlueScript SK （+）,pBlueScript SK（-）pLLP ompA, pINIIIompA, pMBP-P ,pMBP-C, 大肠杆菌冷激质粒： pColdI pColdII pColdIII pColdTF 原核共表达质粒：pACYCduet-1,pETduet-1,pCDFduet-1，pRSFduet-1 Takara公司大肠杆菌分子伴侣： pG-KJE8 pGro7 pKJE7 pGTf2 pTf16 大肠杆菌宿主细胞： DH5a JM101 JM103JM105 JM107 JM109 JM110 Top10 Top10F BL21（DE3）HB101 ER2529 E2566 C2566 MG1655 XL-10gold XL blue M15 JF1125 K802 SG1117 BL21（AI）BL21（DE3）plysS TG1 TB1 DH5a（pir）Tuner（DE3）Bl21 codonplusRIPL Novablue （DE3）Rosetta Rosetta（DE3）Rosetta（DE3）plys Rosetta-gami（DE3）Rosetta-gamiB（DE3）, Rosetta-gamiB（DE3）plysS Orgami（DE3）OrgamiB（DE3）HMS174（DE3）植物表达/RNAi载体农杆菌pBI121,pBI121-GFP,pBI101,pBI221,pSN1301,pUN1301,pRTL2 , pRTL2-GFP , pRTL2-CFP, pRTL2-RFP , pRTL2-YFP,pCAMBIA 1300, 1301, 1302,1303,1304,1305, 1381Z,1391Z,2300, 2301,3300,3301,pCAMBIA super1300,pCAMBIAsuper1300-GFP,pPZP212,pPZP2121,pPZP212-GFP,pGDG,RNAi载体pART27,pHANNIBAL,pKANNIBAL, pFGC5941,pTCK303, pTRV1,pTRV2,T-DNA插入载体（随机突变体库）pSKI015,pSKI074,真菌ATMT载体pBIG2RHPH2-GUS-GFP,pBHt1枯草芽孢杆菌表达载体pWB980,pHT43,pHP13,pHP43,pBE2,pMUTIN4,pUB110,pE194,pMA5, pMK3,pMK4,pHT304,pHY300PLK,pBest502,pDG1363,pSG1154,pAX01, pSAS144,pDL,pDG148-stu,pDG641,pAL12,pUCX05-bgaB,pHT01，配套菌株BS 168,WB600,WB800,WB700,WB800N,1012,FZB42,1A747,广宿主质粒pVLT33RNAi基因沉默干扰敲除载体pSilencer1.0，pSilencer 2.1-U6 hygro， pSilencer 3.1-H1 hygro，pSilencer 3.1-H1 neo， pSilencer 4.1-CMV neo， pSilencer 4.1-CMV puro pMIR-REPORT Luciferase RNAi载体（oligoengine）pSuper-puro RNAi逆转录病毒载体（clontech）: RNAi-Ready pSIREN-Retro Q， RNAi-ReadypSIREN-RetroQ-ZsGreen（Luciferase shRNA Annealed Oligonucleotide）RNAi慢病毒载体（addgene）: pLKO.1哺乳动物表达载体pcDNA3.1+/-,pcDNA4/HisMax B，pSecTag2 A，pVAX1，pBudCE4.1,pTracer CMV2，pcDNA3.1（-）/myc-His A ，pcDNA6-Myc/His B，pCEP4， pIRES，pIRESneo，pIRES hyg3，pCMV-myc，pCMV-HA，pIRES-puro3，pIRES-neo3,pCAGGS哺乳动物双杂交系统pACT，pBIND，pACT-MyoD，pBIND-Id，pG5luc,pCMV-BD, pCMV-AD, pBD-p53, pFR-luc,Cytotrap Two-Hybrid System:pSos, pSos MAFB, pMyr蜕皮激素诱导系统pIND, pVgRxR,LacSwith II哺乳动物诱导表达系统:pOPRSVI ，pOPI3CAT，pCMVLacI,GeneSwitch System:pSwitch哺乳动物表面展示系统:pDisplay, 四环素调控系统（Invitrogen）：pcDNA4/TO/Myc-His A，pcDNA4/TO/Myc-His B，pcDNA4/TO/Myc-His C，pcDNA4/TO/Myc-His/LacZ，pcDNA6/TR四环素调控系统（Clontech）：pTet-On，pTet-Off，pTRE2,pRevTRE，pRevTet-On，pRevTet-off信号通路报告载体:pGAS-TA-Luc，pSTAT3-TA-Luc, pISRE-TA-Luc, pTA-Luc，pIκB-EGFP，pNFAT-TA-Luc，pCaspase3-sensor，pAP1（PMA）-Luc;pGL4.26[luc2P/minP/Hygro],pGL4.29[luc2P/CRE/Hygro],pGL4.30[luc2P/NFA T-RE/Hygro]，pGL4.75;p53-Luc，pAP-1-Luc， pNF-κB-Luc，pSRE-Luc，pFA2-Elk1，pFC-MEKK，pFR-luc,Gateway系统（invitrogen）pcDNA6.2-GWEmGFP-miR negative, pLenti 6/TR,pcDNA 6.2-GW EmGFP-miR，乳酸菌表达载体及各种乳酸菌乳酸杆菌菌株，pNZ8148,pLEISS,pMG36e,pBBR1MCS-5,pBBR1MCS-6,pRV610,pLEM415,pHY3 00PLK,分泌型乳酸菌表达载体pVE5523，pPG611.1,pPG612.1等和乳酸杆菌菌株宿主菌NZ9000,MG1363,Lactobacillus casei 1.539,Lactobacillus casei,acidophilus NCFM,1.2,Lactobacillus sakei 23K,L.plantarum,L.rhamnosusGG,B.coagulans,Bifidobacterium bifidum,Bifidobacterium infantis,Lactococcus lactis M17,1663,Lactobacillus reuterii广宿主表达载体链球菌表达敲除载体假单胞菌表达载体pVLT33,pBBR1MCS-2,3,4,5,6, pJRD215,pJN105,pME6032,Cos载体pLAFR3,pMP2444（GFP）, pHY300PLK,pRT102,pRL1063a, 转座子载体pUT-miniTn5,pMGS100, pWHM10,pKC1139,pSET152,pOJ260,pPG611.1,pPG612.1，腺病毒载体/慢病毒，逆转录病毒表达载体及包装包膜质粒，腺病毒系统（Stratagene）: pAdEasy-1,pShuttle-CMV,pShuttle,pAdTrack, pAdTrack-CMV, pShuttle-IRES-hrGFP-1、pShuttle-IRES-hrGFP-2、pShuttle-CMV-lacZ,pShuttle-CMV-EGFP-C,pXC1, pBHGE3, 配套大肠杆菌BJ5183,293,293T cellline 腺相关病毒系统（Stratagene）： pAAV-MCS，pAAV-RC，pHelper，pAAV-LacZ，pAAV-IRES-hrGFP，pCMV-MCS，慢病毒载体:pLVX-DsRed-Monomer-N1,pLVX-IRES-ZsGreen1,pLVX-AcGFP1-N1,Lenti6/v 5-EDST-EGFP,pWPXL, FUGW,pLentilox 3.7,RNAi-Ready pSIREN-Retro Q,RNAi-Ready pSIREN-Retro Q-ZsGreen,pSUPER.Retro-GFP/Neo,pSUPER-Retro-Neo, pSUPER.Retro-puro,PLNCX PLNCX2 pMSCV-HYG pMSCV-neo pMSCV-puro pLEGFP-C1 pLOX-CW-CRE pLOX-GFP-IRES-TK pRetroX-IRES-DsRedExpress, pLVX-IRES-mCherry质粒载体。

(完整版)所有质粒载体汇总,推荐文档

酿酒酵母表达载体pYES2,pYES2/NT,pYES2/CT,pYES3,pYES6, pYCplac22-GFP,酵母载体pAUR123,pRS303TEF,pRS304, pRS305,pRS306,pY13TEF,pY14TEF，pY15TEF，pY16TEF,酵母基因重组表达载体pUG6, pSH47,酵母单杂载体pHISi,pLacZi,pHIS2, pGAD424, 酵母双杂交系统:酿酒酵母Y187, 酿酒酵母AH109；质粒pGADT7,pGBKT7；对照质粒pGBKT7-53，pGBKT7-lam，pGADT7-T，PCL1，酿酒酵母菌株INVSc1,YM4271, AH109,Y187,Y190,毕赤酵母表达载体pPIC9K,pPIC9K-His,pPIC3.5K,pPICZalphaA,B,C,pPICZA,B,C,pGAPZαA,pAO815,pPIC9k-His,pHIL-S1,pPink hc，配套毕赤酵母Pichiapink,毕赤酵母宿主X33，KM71，KM71H，GS115，原核表达载体pQE30,31,32,40,60,61,62,等原核表达载体,包括pET系列，pET-GST，pGEX系列（含GST标签），pMAL系列pMAL-c2x,-c4x,-c4e,-c5x,-p5x,pBAD,pBADHis,pBADmycHis系列，pQE系列,pTrc99a,pTrcHis系列，pBV220,221,222,pTXB系列，pLLP-ompA,pIN-III-ompA （分泌型表达系列）,pQBI63（原核表达带荧光）pET3a, pET 3d, pET 11a, pET 12a, pET 14b, pET 15b, pET 16b, pET 17b, pET 19b, pET 20b, pET 21a,b,d, pET 22b, pET 23a, pET 23b, pET 24a,b, pET 25b, pET 26b, pET 27b, pET 28a,b, pET 29a, pET 30a, pET 31b, pET 32a, pET 35b, pET 38b, pET 39b, pET 40b, pET 41a,b pET 42a, pET 43.1a,b pET 44a, pET 49b pET302,303 pET His,pET Dsb,pET GST,pET Trx pQE2, pQE9 pQE30,31,32, pQE 40 pQE70 pQE80L pQETirs system pRSET-A pRSET-B pRSET-C pGEX4T-1,-2,-3,5x-1,6p-1,6p-2,2tk,3c pBV220,221,222 pTrcHisA,B,C pBAD24,34,43 pBAD HisA,B,C pPinPoint-Xa1,Xa2,Xa3 pMALc2x, p2x pBV220 pGEM Ex1, pGEM7ZF（+）, pTrc99A, pTwin1, pEZZ18 pkk232-8,pkk233-3,pACYC184,pBR322,pUC119 pTYB1,pTYB2,pTYB4,pTYB11 pBlueScript SK （+）,pBlueScript SK（-）pLLP ompA, pINIIIompA, pMBP-P ,pMBP-C, 大肠杆菌冷激质粒： pColdI pColdII pColdIII pColdTF 原核共表达质粒：pACYCduet-1,pETduet-1,pCDFduet-1，pRSFduet-1 Takara公司大肠杆菌分子伴侣： pG-KJE8 pGro7 pKJE7 pGTf2 pTf16 大肠杆菌宿主细胞： DH5a JM101 JM103JM105 JM107 JM109 JM110 Top10 Top10F BL21（DE3）HB101 ER2529 E2566 C2566 MG1655 XL-10gold XL blue M15 JF1125 K802 SG1117 BL21（AI）BL21（DE3）plysS TG1 TB1 DH5a（pir）Tuner（DE3）Bl21 codonplusRIPL Novablue （DE3）Rosetta Rosetta（DE3）Rosetta（DE3）plys Rosetta-gami（DE3）Rosetta-gamiB（DE3）, Rosetta-gamiB（DE3）plysS Orgami（DE3）OrgamiB（DE3）HMS174（DE3）植物表达/RNAi载体农杆菌pBI121,pBI121-GFP,pBI101,pBI221,pSN1301,pUN1301,pRTL2 , pRTL2-GFP , pRTL2-CFP, pRTL2-RFP , pRTL2-YFP,pCAMBIA 1300, 1301, 1302,1303,1304,1305, 1381Z,1391Z,2300, 2301,3300,3301,pCAMBIA super1300,pCAMBIAsuper1300-GFP,pPZP212,pPZP2121,pPZP212-GFP,pGDG,RNAi载体pART27,pHANNIBAL,pKANNIBAL, pFGC5941,pTCK303, pTRV1,pTRV2,T-DNA插入载体（随机突变体库）pSKI015,pSKI074,真菌ATMT载体pBIG2RHPH2-GUS-GFP,pBHt1枯草芽孢杆菌表达载体pWB980,pHT43,pHP13,pHP43,pBE2,pMUTIN4,pUB110,pE194,pMA5, pMK3,pMK4,pHT304,pHY300PLK,pBest502,pDG1363,pSG1154,pAX01, pSAS144,pDL,pDG148-stu,pDG641,pAL12,pUCX05-bgaB,pHT01，配套菌株BS 168,WB600,WB800,WB700,WB800N,1012,FZB42,1A747,广宿主质粒pVLT33RNAi基因沉默干扰敲除载体pSilencer1.0，pSilencer 2.1-U6 hygro， pSilencer 3.1-H1 hygro，pSilencer 3.1-H1 neo， pSilencer 4.1-CMV neo， pSilencer 4.1-CMV puro pMIR-REPORT Luciferase RNAi载体（oligoengine）pSuper-puro RNAi逆转录病毒载体（clontech）: RNAi-Ready pSIREN-Retro Q， RNAi-ReadypSIREN-RetroQ-ZsGreen（Luciferase shRNA Annealed Oligonucleotide）RNAi慢病毒载体（addgene）: pLKO.1哺乳动物表达载体pcDNA3.1+/-,pcDNA4/HisMax B，pSecTag2 A，pVAX1，pBudCE4.1,pTracer CMV2，pcDNA3.1（-）/myc-His A ，pcDNA6-Myc/His B，pCEP4， pIRES，pIRESneo，pIRES hyg3，pCMV-myc，pCMV-HA，pIRES-puro3，pIRES-neo3,pCAGGS哺乳动物双杂交系统pACT，pBIND，pACT-MyoD，pBIND-Id，pG5luc,pCMV-BD, pCMV-AD, pBD-p53, pFR-luc,Cytotrap Two-Hybrid System:pSos, pSos MAFB, pMyr蜕皮激素诱导系统pIND, pVgRxR,LacSwith II哺乳动物诱导表达系统:pOPRSVI ，pOPI3CAT，pCMVLacI,GeneSwitch System:pSwitch哺乳动物表面展示系统:pDisplay, 四环素调控系统（Invitrogen）：pcDNA4/TO/Myc-His A，pcDNA4/TO/Myc-His B，pcDNA4/TO/Myc-His C，pcDNA4/TO/Myc-His/LacZ，pcDNA6/TR四环素调控系统（Clontech）：pTet-On，pTet-Off，pTRE2,pRevTRE，pRevTet-On，pRevTet-off信号通路报告载体:pGAS-TA-Luc，pSTAT3-TA-Luc, pISRE-TA-Luc, pTA-Luc，pIκB-EGFP，pNFAT-TA-Luc，pCaspase3-sensor，pAP1（PMA）-Luc;pGL4.26[luc2P/minP/Hygro],pGL4.29[luc2P/CRE/Hygro],pGL4.30[luc2P/NFA T-RE/Hygro]，pGL4.75;p53-Luc，pAP-1-Luc， pNF-κB-Luc，pSRE-Luc，pFA2-Elk1，pFC-MEKK，pFR-luc,Gateway系统（invitrogen）pcDNA6.2-GWEmGFP-miR negative, pLenti 6/TR,pcDNA 6.2-GW EmGFP-miR，乳酸菌表达载体及各种乳酸菌乳酸杆菌菌株，pNZ8148,pLEISS,pMG36e,pBBR1MCS-5,pBBR1MCS-6,pRV610,pLEM415,pHY3 00PLK,分泌型乳酸菌表达载体pVE5523，pPG611.1,pPG612.1等和乳酸杆菌菌株宿主菌NZ9000,MG1363,Lactobacillus casei 1.539,Lactobacillus casei,acidophilus NCFM,1.2,Lactobacillus sakei 23K,L.plantarum,L.rhamnosusGG,B.coagulans,Bifidobacterium bifidum,Bifidobacterium infantis,Lactococcus lactis M17,1663,Lactobacillus reuterii广宿主表达载体链球菌表达敲除载体假单胞菌表达载体pVLT33,pBBR1MCS-2,3,4,5,6, pJRD215,pJN105,pME6032,Cos载体pLAFR3,pMP2444（GFP）, pHY300PLK,pRT102,pRL1063a, 转座子载体pUT-miniTn5,pMGS100, pWHM10,pKC1139,pSET152,pOJ260,pPG611.1,pPG612.1，腺病毒载体/慢病毒，逆转录病毒表达载体及包装包膜质粒，腺病毒系统（Stratagene）: pAdEasy-1,pShuttle-CMV,pShuttle,pAdTrack, pAdTrack-CMV, pShuttle-IRES-hrGFP-1、pShuttle-IRES-hrGFP-2、pShuttle-CMV-lacZ,pShuttle-CMV-EGFP-C,pXC1, pBHGE3, 配套大肠杆菌BJ5183,293,293T cellline 腺相关病毒系统（Stratagene）： pAAV-MCS，pAAV-RC，pHelper，pAAV-LacZ，pAAV-IRES-hrGFP，pCMV-MCS，慢病毒载体:pLVX-DsRed-Monomer-N1,pLVX-IRES-ZsGreen1,pLVX-AcGFP1-N1,Lenti6/v 5-EDST-EGFP,pWPXL, FUGW,pLentilox 3.7,RNAi-Ready pSIREN-Retro Q,RNAi-Ready pSIREN-Retro Q-ZsGreen,pSUPER.Retro-GFP/Neo,pSUPER-Retro-Neo, pSUPER.Retro-puro,PLNCX PLNCX2 pMSCV-HYG pMSCV-neo pMSCV-puro pLEGFP-C1 pLOX-CW-CRE pLOX-GFP-IRES-TK pRetroX-IRES-DsRedExpress, pLVX-IRES-mCherry质粒载体。

维萨拉验证服务商品说明书

维萨拉验证服务/ 高效、合规的温湿度分布试验卓越的质量管理体系为确保满足GM P 《药品生产质量管理规范》的要求，各机构必须证明其仓储和运输区域的环境条件都处于适当的控制下。

因此，质量计划的首要步骤应是通过分布研究深入了解这些区域的温度属性。

然而，分布研究需要的不仅仅是设备,还需要验证和校准方面的专业知识与技能。

维萨拉为您提供由娴熟的技术人员完成的世界级验证服务，服务包含用以进行分布研究，内容翔实的验证计划及分布试验协议。

深入透彻的验证后报告将会提升您对受控环境的了解。

这些文档和专业技能将有助于体现您的企业正在遵循受控环境验证与文档记录的当前最佳实践。

我们提供验证所需的专业技能和设备服务，所以您的质量和设施经理能专注于他们的日常工作。

降低生命科学环境的风险在过去20年间，我们已为生命科学行业的受控环境专门设计了创新型设备和软件，这些行业包括：• 制药• 生物技术• 医疗器械• 营养和保健品应用领域：• 冰箱/冷冻柜/冷藏室 • 培养箱• 稳定性试验箱• 仓库/存储区/批发中心翔实、精确的存档为进行分布研究，验证服务技术人员将会收集并分析您所在区域、房间或试验箱的数据，辨识工艺过程中的热点或冷点。

技术人员可对空载/满载试验箱，或仓储区的静态/动态过程进行挑战性实验。

测试可能会根据应用需要部署热电偶或配有传感器的数据记录仪，使用维萨拉验证软件下载分析测试数据。

所有分布研究文档均为安全、演示级品质。

报告包含最小值、最大值以及平均值统计和平均动力学温度(MKT) 数据,可轻松导出为供深入分析使用的通用.csv文件。

温度传感器工作范围：-90 °C至70 °C，精度高达±0.1 °C，分辨率0.02 °C.相对湿度传感器工作范围：10%至90%，精度高达±1%，分辨率0.05%.合规性： 21 CFR Part 11， E U Annex 11 21随着全球范围的监管机构对制药、生物技术和医疗器械制造商和经销商的审查日趋严格，许多企业开始为其受控环境寻求验证解决方案。

辅助肥皂和脂肪吸收器泡沫类产品说明书

UnitedHealthcare PharmacyClinical Pharmacy ProgramsProgram Number 2023 P 1114-12Program Prior Authorization/NotificationMedication Includes both brand and generic versions and all formulations ofthe listed products unless otherwise noted: Weight Loss –phentermine (all brand products including Adipex-P and Lomaira),benzphetamine, Contrave (naltrexone HCl and bupropion HCl,diethylpropion, Imcivree (setmelanotide), phendimetrazine, orlistat(Xenical), Qsymia (phentermine and topiramate extended-release),Saxenda (liraglutide), and Wegovy (semaglutide)P&T Approval Date 1/08, 1/2010, 1/2011, 1/2012, 11/2012, 11/2013, 11/2014, 7/2015,9/2016, 9/2017, 9/2018, 9/2019, 9/2020, 4/2021, 9/2021, 9/2022,2/2023Effective Date 5/1/2023;Oxford: 5/1/20231.Background:This is an optional program that is put in place for clients or businesses that have elected to cover weight loss products with Prior Authorization/Notification. It is also designed to meet regulatory requirements for coverage of weight loss medications in California, Maryland, New Mexico and New York.Classification BMI(kg/m2)Underweight< 18.50Normal range 18.50 - 24.99Overweight ≥ 25.00Obese ≥ 30.00Obese class I 30.00 - 34.99Obese class II 35.00 - 39.99Obese class III ≥ 40.00aa State mandates may apply. Any federal regulatory requirements and the member specificbenefit plan coverage may also impact coverage criteria. Other policies and utilizationmanagement programs may apply.3.Additional Clinical Rules:•Supply limits may be in place.4.References:1. American Association of Clinical Endocrinologists and American College ofEndocrinology Comprehensive Clinical Practice Guidelines for Medical Care ofPatients with Obesity. Endocr Pract. 2016;22(Suppl 3).2. National Institutes of Health, National Heart, Lung, and Blood Institute, and NorthAmerican Association for the Study of Obesity. The Practical Guide: Identification,Evaluation, and Treatment of Overweight and Obesity in Adults. 2000.3. American Gastroenterological Association medical position statement on obesity.Gastroenterology 2002 Sep;123(3):879-81.4. Bray, GA, Ryan, DH. Medical Therapy for the Patient with Obesity. Circulation.2012;125:1695-1703.5. Barlow, SE. Expert Committee Recommendations Regarding the Prevention, Assessment,and Treatment of Child and Adolescent Overweight and Obesity: Summary Report.Pediatrics 2007.6. Benzphetamine [package insert]. Laurelton, NY: Epic Pharma, LLC; January 2021.7. Diethylpropion [package insert]. Philadelphia, PA: Lannett Company, Inc.; December2019.8. Phendimetrazine Slow-Release Capsules [package insert]. Princeton, NJ: Sandoz Inc.;November 2018.9. Adipex-P [package insert]. Parsippany, NJ: Teva Pharmaceuticals USA, Inc; s ;March2020. 10. X enical [package insert]. South San Franciso, CA : Roche Pharmaceuticals;September 2020.10. Qsymia [package insert]. Moutain View, CA. Vivus, Inc ; June 2022.11 Contrave [package insert]. Brentwood, TN: Currax Pharmaceuticals, LLC; November2021.12. Saxenda [package insert]. Plainsboro, NJ: Novo Nordisk ; June 2022.13. Pharmacological Management of Obesity: An Endocrine Society Clinical PracticeGuideline. The Journal of Clinical Endocrinology & Metabolism 2015 100:2, 342-36214. AHA/ACC/TOS Prevention Guideline: 2013 AHA/ACC/TOS Guideline for theManagement of Overweight and Obesity in Adults: A Report to the American College ofCardiology/American Health Association Task Force on Practice Guidelines and TheObesity Society. Circulation. 2014; 129:S102-13815. World Health Organization. (2006). Global Database on Body Mass Index. Retrievedfrom https://www.who.int/data/gho/data/themes/topics/topic-details/GHO/body-mass-index16. Lomaira [package insert]. Newtown, PA: KVK-Tech, Inc. December 2018.17. Imcivree [package insert]. Boston, MA: Rhythm Pharmaceuticals, Inc; June 2022.18. Wegovy [package insert]. Plainsboro, NJ: Novo Nordisk; December 2022.Program Weight LossChange ControlDate Change11/2013 Reformatted to standard and updated references. Clarified risk factorsrequired in addition to BMI of 27-29. Removed criteria that coveragewould not be provided if used in combination with other weight lossagents. Shortened initial authorization for sympathomimetics to 3months based on PI recommendations for short-term use. Addedreauthorization criteria for 6 months, changed initial authorizationperiod of Xenical from 4 months to 3 months, and shortenedreauthorization of Belviq and Xenical to 12 months for alignment withOptumRx.11/2014 Added Contrave to guideline.7/2015 Added Saxenda to guideline.9/2016 Annual Review. Added Belviq XR to guideline. Revised authorizationperiod for Xenical. Removed ASO from title- program will remain asASO option.9/2017 Annual Review. Added Lomaira. Updated references. Revisedweightloss percentage for Saxenda per PI.9/2018 Annual Review. Updated references. .9/2019 Annual Review. Revised background section. Revised list ofmodifications of weight related comorbidity.9/2020 Annual review. Updated references. Removed Belviq/Belviq XR.4/2021 Added Imcivree as in scope. Added Imcivree criteria. Updatedreferences. Formatting changes.9/2021 Added Wegovy to criteria. Updated Saxenda criteria to allow forcoverage for 12 years and older. Updated Qsymia weight loss goal togreater than 3 percent per label and changed initial authorization to 4months. Updated references.9/2022 Added background note that this program is intended to meet regulatoryrequirements for coverage of weight loss medications in California,Maryland, New Mexico and New York. This program is replacingpolicy P 1172-8. Changed Qsymia age requirement to 12 or older due tonew pediatric labeling. Updated Imcivree to include new FDAapproved indication for Bardet-Biedl syndrome. Updated references. 2/2023 Updated Wegovy criteria to include pediatric patients based on newlabeling. Added state mandate language.。

(完整版)所有质粒载体汇总,推荐文档

酿酒酵母表达载体pYES2,pYES2/NT,pYES2/CT,pYES3,pYES6, pYCplac22-GFP,酵母载体pAUR123,pRS303TEF,pRS304, pRS305,pRS306,pY13TEF,pY14TEF，pY15TEF，pY16TEF,酵母基因重组表达载体pUG6, pSH47,酵母单杂载体pHISi,pLacZi,pHIS2, pGAD424, 酵母双杂交系统:酿酒酵母Y187, 酿酒酵母AH109；质粒pGADT7,pGBKT7；对照质粒pGBKT7-53，pGBKT7-lam，pGADT7-T，PCL1，酿酒酵母菌株INVSc1,YM4271, AH109,Y187,Y190,毕赤酵母表达载体pPIC9K,pPIC9K-His,pPIC3.5K,pPICZalphaA,B,C,pPICZA,B,C,pGAPZαA,pAO815,pPIC9k-His,pHIL-S1,pPink hc，配套毕赤酵母Pichiapink,毕赤酵母宿主X33，KM71，KM71H，GS115，原核表达载体pQE30,31,32,40,60,61,62,等原核表达载体,包括pET系列，pET-GST，pGEX系列（含GST标签），pMAL系列pMAL-c2x,-c4x,-c4e,-c5x,-p5x,pBAD,pBADHis,pBADmycHis系列，pQE系列,pTrc99a,pTrcHis系列，pBV220,221,222,pTXB系列，pLLP-ompA,pIN-III-ompA （分泌型表达系列）,pQBI63（原核表达带荧光）pET3a, pET 3d, pET 11a, pET 12a, pET 14b, pET 15b, pET 16b, pET 17b, pET 19b, pET 20b, pET 21a,b,d, pET 22b, pET 23a, pET 23b, pET 24a,b, pET 25b, pET 26b, pET 27b, pET 28a,b, pET 29a, pET 30a, pET 31b, pET 32a, pET 35b, pET 38b, pET 39b, pET 40b, pET 41a,b pET 42a, pET 43.1a,b pET 44a, pET 49b pET302,303 pET His,pET Dsb,pET GST,pET Trx pQE2, pQE9 pQE30,31,32, pQE 40 pQE70 pQE80L pQETirs system pRSET-A pRSET-B pRSET-C pGEX4T-1,-2,-3,5x-1,6p-1,6p-2,2tk,3c pBV220,221,222 pTrcHisA,B,C pBAD24,34,43 pBAD HisA,B,C pPinPoint-Xa1,Xa2,Xa3 pMALc2x, p2x pBV220 pGEM Ex1, pGEM7ZF（+）, pTrc99A, pTwin1, pEZZ18 pkk232-8,pkk233-3,pACYC184,pBR322,pUC119 pTYB1,pTYB2,pTYB4,pTYB11 pBlueScript SK （+）,pBlueScript SK（-）pLLP ompA, pINIIIompA, pMBP-P ,pMBP-C, 大肠杆菌冷激质粒： pColdI pColdII pColdIII pColdTF 原核共表达质粒：pACYCduet-1,pETduet-1,pCDFduet-1，pRSFduet-1 Takara公司大肠杆菌分子伴侣： pG-KJE8 pGro7 pKJE7 pGTf2 pTf16 大肠杆菌宿主细胞： DH5a JM101 JM103JM105 JM107 JM109 JM110 Top10 Top10F BL21（DE3）HB101 ER2529 E2566 C2566 MG1655 XL-10gold XL blue M15 JF1125 K802 SG1117 BL21（AI）BL21（DE3）plysS TG1 TB1 DH5a（pir）Tuner（DE3）Bl21 codonplusRIPL Novablue （DE3）Rosetta Rosetta（DE3）Rosetta（DE3）plys Rosetta-gami（DE3）Rosetta-gamiB（DE3）, Rosetta-gamiB（DE3）plysS Orgami（DE3）OrgamiB（DE3）HMS174（DE3）植物表达/RNAi载体农杆菌pBI121,pBI121-GFP,pBI101,pBI221,pSN1301,pUN1301,pRTL2 , pRTL2-GFP , pRTL2-CFP, pRTL2-RFP , pRTL2-YFP,pCAMBIA 1300, 1301, 1302,1303,1304,1305, 1381Z,1391Z,2300, 2301,3300,3301,pCAMBIA super1300,pCAMBIAsuper1300-GFP,pPZP212,pPZP2121,pPZP212-GFP,pGDG,RNAi载体pART27,pHANNIBAL,pKANNIBAL, pFGC5941,pTCK303, pTRV1,pTRV2,T-DNA插入载体（随机突变体库）pSKI015,pSKI074,真菌ATMT载体pBIG2RHPH2-GUS-GFP,pBHt1枯草芽孢杆菌表达载体pWB980,pHT43,pHP13,pHP43,pBE2,pMUTIN4,pUB110,pE194,pMA5, pMK3,pMK4,pHT304,pHY300PLK,pBest502,pDG1363,pSG1154,pAX01, pSAS144,pDL,pDG148-stu,pDG641,pAL12,pUCX05-bgaB,pHT01，配套菌株BS 168,WB600,WB800,WB700,WB800N,1012,FZB42,1A747,广宿主质粒pVLT33RNAi基因沉默干扰敲除载体pSilencer1.0，pSilencer 2.1-U6 hygro， pSilencer 3.1-H1 hygro，pSilencer 3.1-H1 neo， pSilencer 4.1-CMV neo， pSilencer 4.1-CMV puro pMIR-REPORT Luciferase RNAi载体（oligoengine）pSuper-puro RNAi逆转录病毒载体（clontech）: RNAi-Ready pSIREN-Retro Q， RNAi-ReadypSIREN-RetroQ-ZsGreen（Luciferase shRNA Annealed Oligonucleotide）RNAi慢病毒载体（addgene）: pLKO.1哺乳动物表达载体pcDNA3.1+/-,pcDNA4/HisMax B，pSecTag2 A，pVAX1，pBudCE4.1,pTracer CMV2，pcDNA3.1（-）/myc-His A ，pcDNA6-Myc/His B，pCEP4， pIRES，pIRESneo，pIRES hyg3，pCMV-myc，pCMV-HA，pIRES-puro3，pIRES-neo3,pCAGGS哺乳动物双杂交系统pACT，pBIND，pACT-MyoD，pBIND-Id，pG5luc,pCMV-BD, pCMV-AD, pBD-p53, pFR-luc,Cytotrap Two-Hybrid System:pSos, pSos MAFB, pMyr蜕皮激素诱导系统pIND, pVgRxR,LacSwith II哺乳动物诱导表达系统:pOPRSVI ，pOPI3CAT，pCMVLacI,GeneSwitch System:pSwitch哺乳动物表面展示系统:pDisplay, 四环素调控系统（Invitrogen）：pcDNA4/TO/Myc-His A，pcDNA4/TO/Myc-His B，pcDNA4/TO/Myc-His C，pcDNA4/TO/Myc-His/LacZ，pcDNA6/TR四环素调控系统（Clontech）：pTet-On，pTet-Off，pTRE2,pRevTRE，pRevTet-On，pRevTet-off信号通路报告载体:pGAS-TA-Luc，pSTAT3-TA-Luc, pISRE-TA-Luc, pTA-Luc，pIκB-EGFP，pNFAT-TA-Luc，pCaspase3-sensor，pAP1（PMA）-Luc;pGL4.26[luc2P/minP/Hygro],pGL4.29[luc2P/CRE/Hygro],pGL4.30[luc2P/NFA T-RE/Hygro]，pGL4.75;p53-Luc，pAP-1-Luc， pNF-κB-Luc，pSRE-Luc，pFA2-Elk1，pFC-MEKK，pFR-luc,Gateway系统（invitrogen）pcDNA6.2-GWEmGFP-miR negative, pLenti 6/TR,pcDNA 6.2-GW EmGFP-miR，乳酸菌表达载体及各种乳酸菌乳酸杆菌菌株，pNZ8148,pLEISS,pMG36e,pBBR1MCS-5,pBBR1MCS-6,pRV610,pLEM415,pHY3 00PLK,分泌型乳酸菌表达载体pVE5523，pPG611.1,pPG612.1等和乳酸杆菌菌株宿主菌NZ9000,MG1363,Lactobacillus casei 1.539,Lactobacillus casei,acidophilus NCFM,1.2,Lactobacillus sakei 23K,L.plantarum,L.rhamnosusGG,B.coagulans,Bifidobacterium bifidum,Bifidobacterium infantis,Lactococcus lactis M17,1663,Lactobacillus reuterii广宿主表达载体链球菌表达敲除载体假单胞菌表达载体pVLT33,pBBR1MCS-2,3,4,5,6, pJRD215,pJN105,pME6032,Cos载体pLAFR3,pMP2444（GFP）, pHY300PLK,pRT102,pRL1063a, 转座子载体pUT-miniTn5,pMGS100, pWHM10,pKC1139,pSET152,pOJ260,pPG611.1,pPG612.1，腺病毒载体/慢病毒，逆转录病毒表达载体及包装包膜质粒，腺病毒系统（Stratagene）: pAdEasy-1,pShuttle-CMV,pShuttle,pAdTrack, pAdTrack-CMV, pShuttle-IRES-hrGFP-1、pShuttle-IRES-hrGFP-2、pShuttle-CMV-lacZ,pShuttle-CMV-EGFP-C,pXC1, pBHGE3, 配套大肠杆菌BJ5183,293,293T cellline 腺相关病毒系统（Stratagene）： pAAV-MCS，pAAV-RC，pHelper，pAAV-LacZ，pAAV-IRES-hrGFP，pCMV-MCS，慢病毒载体:pLVX-DsRed-Monomer-N1,pLVX-IRES-ZsGreen1,pLVX-AcGFP1-N1,Lenti6/v 5-EDST-EGFP,pWPXL, FUGW,pLentilox 3.7,RNAi-Ready pSIREN-Retro Q,RNAi-Ready pSIREN-Retro Q-ZsGreen,pSUPER.Retro-GFP/Neo,pSUPER-Retro-Neo, pSUPER.Retro-puro,PLNCX PLNCX2 pMSCV-HYG pMSCV-neo pMSCV-puro pLEGFP-C1 pLOX-CW-CRE pLOX-GFP-IRES-TK pRetroX-IRES-DsRedExpress, pLVX-IRES-mCherry质粒载体。

Infinity Alpha Center Bois Cl Codic系列产品使用说明说明书

MARQUE: INFINITY REFERENCE: ALPHA CENTER BOIS CL CODIC: 0323098INSTRUCTION MANUALBEDIENUNGSANLEITUNGMODE D’EMPLOI MANUAL DE INSTRUCCIONES GEBRUIKSAANWIJZINGMANUALE D’ISTRUZIONIBRUGSVEJLEDNING MALLISTON KÄYTTÖOHJE MANUAL DE INSTRUÇÕESMode d’emploi des enceintes Série ALPHA InfinityNous vous félicitons pour votre choix des enceintes Série ALPHA Infinity. Les instructions contenues dans ce manuel vous aideront à tirer le meilleur parti de ces enceintes.EMBALLAGELe conditionnement de vos enceintes Série ALPHA a été réalisé pour les protéger de tout dommage qui pourrait résulter d’une manutention peu délicate ou du transport. Nous vous conseillons vivement de les conserver pour les déménagements ultérieurs ou pour d’éventuelles opérations de maintenance.POSITIONNEMENTUn positionnement correct des enceintes au sein de la pièce est de première importance pour obtenir le meilleur son possible. Les réflexions sur des murs réverbérants, l’emplacement du mobilier, l’existence de tentures ou rideaux épais, la présence de baies vitrées etc. peuvent influencer notablement la qualité du son, ou l’équilibre et la bonne restitution de l’image stéréo. Il est essentiel d’essayer plusieurs solutions pour déterminer quel positionnement donne la meilleure balance générale et la meilleure image sonore. Pour un effet stéréo optimum les enceintes devraient êtres distantes d’au moins 2 à 3 mètres. Parfois, le fait de les tourner légèrement àl’intérieur, vers l’auditeur, augmente la spatialisation et donne une représentation sonore plus exacte. Évitez, autant que possible, de placer ces enceintes contre les murs ou dans les angles, qui engendrent une image stéréo médiocre et un excès de graves. Si vous devez, pour des raisons techniques, placer les haut-parleurs au dessus du niveau des oreilles, inclinez-les vers le bas pour diriger les tweeters dans l’axe d’écoute de l’auditeur.INSTALLATION DES POINTES DE DECOUPLAGETous les modèles posés au sol sont équipés de pointes de découplage destinées àisoler l’enceinte des vibrations retransmises par le sol.Pour les installer, suivez les instructions1.R etournez l’enceinte à l’envers,reposée là sur sa partie supérieure.2.Enfilez un écrou au bout de chaquepointes de découplage, jusqu’à cequ’il atteigne la partie non filletée.3.Vissez les pointes de découplagedans les emplacements prévus sur labase de l’enceinte.4.Réglez-les pour qu’elles aient la mêmehauteur, puis dévissez le contre-écrou pour les bloquer et garder ceréglage.7CONNEXIONSVérifiez bien que tous les appareils soient éteints avant d’effectuer les branchements.Pour brancher les enceintes au récepteur ou à l’amplificateur, utilisez un câble isolé, àdeux conducteurs. Votre revendeur Infinity pourra vous conseiller sur les modèles adaptés, mais nous recommandons de toutes façons un diamètre supérieur à 1,3 mm (16 AWG), et même nettement plus si vos enceintes sont distantes de plus de 10 m de leur ampli ou récepteur. Le câblage est une partie importante des systèmes audio.Vous pouvez même légitimement souhaiter faire des essais comparatifs chez votre revendeur de différents ensembles ou types de câbles qu’il propose. Pour chaque voie, la borne rouge de l’enceinte doit être reliée à la borne repérée rouge ou (+) de l’ampli (ou récepteur), et la noire à sa correspondante noire ou (-). De cette manière,les enceintes seront «en phase » et non en opposition. Le fait de brancher les enceintes hors-phase ne les endommagera pas, mais réduira à la fois la séparation stéréo et la qualité des basses. La plupart des conducteurs pour haut-parleurs sont repérés soit par leur couleur, soit par la présence d’une gouttière ou d’une rainure sur le gainage d’un des deux fils. Cela permet d’identifier facilement le fil à connecter àchaque terminal. Si vous voulez branchez plus d’une paire d’enceintes sur un ampli ou un récepteur, vérifiez bien auprès de votre revendeur ou du constructeur de l’ampli qu’il puisse en supporter la charge en toute sécurité.clippent » et génèrent une distorsion qui des conditions limites et dangereuses pour les éléments les plus faibles du système.Plus l’ampli que vous utilisez est petit, et plus grand est le risque d’écréter, et donc de mettre en danger les tweeters. Les réglages graves/aigus de l’ampli ou du récepteur peuvent aussi exagérer ou renforcer ce phénomène. Nous vous conseillons donc de 8toujours utiliser un ampli ou un récepteur susceptible de faire fonctionner vos enceintes ALPHA raisonnablement fort tout en gardant un son clair et non distordu. ENCEINTE CENTRALE ALPHALa série ALPHA d’INFINITY dispose enceinte centrale dédiée. L’ALPHA Center est amagnétique ce qui vous permet de la positionner directement et en toute sécuritésur, ou sous votre téléviseur. La face avant de l'ALPHA Center est conçue pour s’aligner le mieux possible à la forme du téléviseur. L’ALPHA Center se branche sur un ampli/ampli-tuner audio-vidéo de la même manière qu’une enceinte conventionnelle. (le + sur le + et le - sur le -).ENTRETIEN DE VOS ENCEINTESL'ébenisterie de vos enceintes Infinity est recouvert d'un placage vinyl de haute qualité nécessitant peu d'entretien.Dépoussiérez l’ébénisterie avec un chiffon humide ou un produit de nettoyage pour meubles. Les grilles peuvent être passées àl’aspirateur, réglé sur la puissance minimale.EN CAS DE PROBLEMES DANS VOTRE SYSTEME STÉRÉOAvant d’appeller votre revendeur, le service consommateurs d’Infinity ou le distributeur Infinity de votre pays, vous pouvez réaliser quelques tests afin de localiser et résoudre certains problèmes concernant vos enceintes. Vous pouvez utiliser simplement les deux canaux stéréo de votre ampli pour découvrir l’origine de la panne : en cas de son distordu, l’écoute séparée de chaque enceinte permet de savoir si le problème est présent des deux côtés ou d’un seul. Dans le premier cas, le problème provient sans doute d’un autre élément de votre chaîne. S’il ne concerne qu’une voie, essayez d’inverser les sorties de l’ampli. Si la distorsion passe sur l’autre enceinte, làencore, le problème ne vient pas des écoutes mais réside plus en amont dans la chaîne.PROBLEMES CONCERNANT LES ENCEINTESSi une voie (tweeter, médium, boomer) de votre enceinte ne fonctionne pas, retirez le haut parleur correspondant de son logement et vérifiez que les fils sont bien solidaires des cosses du HP.Si ce n’est pas le cas, rebranchez-les et soudez-les correctement. Si les branchements sont corrects mais que le haut parleur ne fonctionne quand même pas, débranchez-le, retirez-le et remplacez-le par son homologue de l’autre enceinte. Si cela résout le problème, installez le haut-parleur suspect dans l’enceinte dont vous avez extrait ce HP de remplacement. S'il ne fonctionne pas mieux, vous pouvez être certain que c’est bien lui qui est défectueux et qu’il faut le remplacer.RÉPARATIONSPour tous problèmes de réparations, rapportez vos enceintes au revendeur chez qui vous avez effectué l’achat. Si, pour une raison ou une autre, cela n’était pas possible, contactez votre distributeur Infinity local.Harman France - Z.A du Parc Terminal - 9 rue St Just -93135 NOISY LE SEC Cedex9。

奥林匹亚产品说明书

PORCELAINAll items shown in this document are part of Olympia’s stocking program. For special orders, please contact your Olympia Tile Sales Representative.30 x 60 cm (12 x 24)Matte RF.PS.STT.1224.MT Polished RF.PS.STT.1224.PL 75 x 75 cm (30 x 30)Polished RF.PS.STT.3030.PL30 x 60 cm (12 x 24)Matte RF.PS.BOT.1224.MT Polished RF.PS.BOT.1224.PL 75 x 75 cm (30 x 30)Polished RF.PS.BOT.3030.PLSTATUARIO (White)BOTTICINOAll items shown in this document are part of Olympia’s stocking program. For special orders, please contact your Olympia Tile Sales Representative.30 x 60 cm (12 x 24)Matte RF.PS.MCB.1224.MT Polished RF.PS.MCB.1224.PL 75 x 75 cm (30 x 30)Polished RF.PS.MCB.3030.PL30 x 60 cm (12 x 24)Matte RF.PS.IPO.1224.MT Polished RF.PS.IPO.1224.PL 75 x 75 cm (30 x 30)Polished RF.PS.IPO.3030.PLMACAUBASIMPEROAll items shown in this document are part of Olympia’s stocking program. For special orders, please contact your Olympia Tile Sales Representative.30 x 60 cm (12 x 24)Matte RF.PS.ACD.1224.MT Polished RF.PS.ACD.1224.PL 75 x 75 cm (30 x 30)Polished RF.PS.ACD.3030.PL30 x 60 cm (12 x 24)Matte RF.PS.PUL.1224.MT Polished RF.PS.PUL.1224.PL 75 x 75 cm (30 x 30)Polished RF.PS.PUL.3030.PLARCADIA (Taupe)PULPIS (Brown)Technical data is supplied by the manufacturer and is subject to change at any time. Olympia does not provide warranties as to thespecifications of any product.Water Absorption ISO 10545-3< 0.5%ConformsPrestigio Series Tests Performed -- Results ConformColour VariationMemberMember of Canada Green BuildingCouncilMember of U.S. Green BuildingCouncilFor information concerning LEED, please contact our Sales RepresentativeExamples of Recommended ApplicationsResidentialCommercialIndoorOutdoorWallFloorDistributors of America (CTDA)The Terrazzo, Tile, and Marble Association ofCanadaMarble Institute of AmericaTaskGrout Sealing MORE™ Grout, Ceramic &Porcelain Sealer Ready to use Commercial: 1-3 Years Residential: 5-8 YearsDaily Maintenance MORE™ Stone and TileCleaner 1 : 128 Ratio Daily Deep Cleaning MORE™ Alkaline Cleaner 1 : 4 Ratio As Needed Grout Haze- CementitiousMORE™ Acidic Cleaner 1 : 4 Ratio As Needed Grout Haze - Epoxy / UrethaneMORE™ Coating Stripper*Ready to useAs Needed* Always test in an inconspicuous area to make sure the product is achieving the desired results and is not damaging the finish of the surface.SURFACE PREPARATION (If Required)• Floor levelling: 59 Flex Flo / 5900 Flex Flo Plus Self Levelling Underlayment • Waterproofing: WP900 Waterproof Membrane• Crack Isolation/Uncoupling: Flexilastic 1000 Crack Isolation Membrane / FlexMat Uncoupling Membrane* For a complete listing of FLEXTILE products and accessories including Sound Control, In‐Floor Heating, and further setting options, please visit ARCADIA (Taupe)MACAUBASIt is Olympia Tile’s policy to stock all items for sale. Should you require additional sizes, finishes or colours, Olympia has a Special Order Department to help find exactly what you are looking for. Please contact your Olympia Tile Sales Representative for more details. Note that minimum order quantities and longer lead times apply to all special order items.Colour may vary slightly from images shown. It is highly recommended to view physical samples of tile prior to purchase. Note that shade variations between tiles is characteristic and adds to the inherent beauty of the product. Please visit a showroom near you or request a sample from your Olympia Tile Sales Representative.Prestigio Series: Board MEF-136 / Folder FL-777。

爱普灵操作指南

在左上方工具条内选择
‹#› ApneaLink Plus © ResMed 2013
‹#› ApneaLink Plus © ResMed 2013
参数设置—低通气
气流强度下降到%
AASM标准
最大持续时间
最小持续时间
两个事件暂停/ 低通气之间的最小时间间隔
在Tool菜单下Seting--Analysis parameters
‹#› ApneaLink Plus © ResMed 2013
软件一
• 点开索引图示，如右图共分三部分 1、病人信息和 ApneaLink准备或下载时输入病人信息一样 2、当大量数据时，可以输入姓、名或生日搜索你需要的信息
病人信息报告趋势图
‹#› ApneaLink Plus © ResMed 2013
软件三---趋势图
1、上界面（窗口）
2、下界面时间段标记 3、时间标记线 4、信号 5、通道 6、下界面（窗口） 7、滚动条或方向指示
8、事件
9、事件类型选择
10、从一个事件到另一个事件的转换
11、选择每屏时间段 12、信号基线调整 13、信us © ResMed 2013
软件四---手动分析
排除分析中枢性暂停
结束分析
气流受限合并鼾声
气流受限
低通气
吸气受限
阻塞性暂停开始分析
混合性暂停
气流强度阈值，下降到%
最少持续时间最大持续时间暂停事件大于60%的时间符合中枢性的，定义为中枢性
气流降低了% 暂停事件小于 20%的时间符合中枢性的，定义为阻塞性呼吸努力度阈值气流强度下降到%
在Tool 菜单下Setting— Analysis parameter