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类志贺邻单胞菌的检测方法概述

类志贺邻单胞菌的检测方法概述

技术推/*2019年第4期n.rr jouknalOF ASKtCULTUItAL aCIBNCB*类志贺邻单胞菌的检测方法概述陈美群,扎西拉姆,潘瑛子(西藏自治区农牧科学院水产科学研究所,西藏拉萨850032)摘要:类志贺邻单胞菌是一种独特的革兰氏阴性、具极端鞭毛的人-兽共患病原菌,该菌可导致许多动物尤其鱼类等水生动物疾病频发,同时可引起腹泻、脑膜炎、败血症、蜂窝组织炎等人类疾病,具有较高的传染性和发病率。

由于该菌不能及时鉴定而导致的延误治疗,不仅给水产养殖业造成重大经济损失,同时也给消费者带来重大安全隐患。

为此快速、准确的鉴别出该病原菌,有针对性的开展药物防治尤为重要。

本文对该菌的传统检测技术,免疫学诊断技术,分子生物学检测技术进行了详细概述,以期为该菌的快速、准确诊断提供详尽资料。

关键词:类志贺邻单胞菌;传统检测;免疫学诊断;分子生物学检测中图分类号:S9643文献标识码:AReaearch Progress of Detection Method of Plesiomonas shigelloidesCHEN Mei-qun,Zhaxilamu,PAN Ying-zi*(Institute of Fisheries Science,Tibet Academy of Agricultural and Animal Husbandry Sciences,Tibet Lhasa850032,China)Abstract:Plesiomonas shigelloides is a unique gram-negative,ultra flagellum human-animal pathogen.P.shigelloides can cause many ani­mals,especially fish and other aquatic animal diseases,and it also can cause diarrhea,meningitis,sepsis,hibritis and other human disea­ses,with high infectious and morbidity rates.Due to the delay of fail to identify P.shigelloides in time,it not only caused major economic losses to aquaculture industry,but also brought major safety hazards to consumers.Rapid and accurate identification of P.shigelloides was particularly important to prevention and control of it.In order to provide detailed information for the rapid and accurate diagnosis of P.shig­elloides,the traditional detection techniques,immunology diagnosis techniques and molecular biology detection techniques were summarized in this paper.Key words:Plesiomonas shigelloides;Traditional detection;Immunological diagnosis;Molecular biological detection类志贺邻单胞菌(Plesiomonas shigelloides))是一种革兰氏阴性、氧化酶阳性的运动性杆菌,隶属肠杆菌科(Enterobacteriaceae)邻单胞菌属(Plesiomonas Habs and Schubert,1962)内唯一的一个种⑴。

自己翻译的罗氏tunel检测细胞凋亡试剂盒说明书

自己翻译的罗氏tunel检测细胞凋亡试剂盒说明书

罗氏tunel检测细胞凋亡试剂盒说明书注意:Label溶液含有甲次砷酸盐和二氯化钴,严禁吸入和食入。

反应悬浮物收集于密闭、不易碎、有明确标识的容器中,按有毒废物处理。

除上表所列试剂外,还需准备以下溶液。

下表列出每步所需物品概览:特异性:TUNEL反应优先标记凋亡产生的DNA链断裂,从而辨别凋亡与坏死、以及由抑制细胞生长的药物或放射线产生的primary DNA链断裂实验干扰:假阴性:在某些型式的凋亡细胞中DNA链断裂可能缺失或不完全。

空间位阻,如细胞外元件可能阻止TdT到达DNA断裂处。

两种情况均能产生假阴性。

假阳性:在坏死晚期,可能产生大量的DNA片段DNA链断裂也可能在具有高增殖和代谢活动的细胞中出现。

两种情况均能产生假阳性。

为确认细胞死亡的凋亡型式,应认真进行每种细胞的形态学检查凋亡过程中产生的形态学改变尤其特征形式,因此,对于可以结果进行解释时,细胞形态评估是一项重要的参数样本:细胞离心涂片和细胞涂片在chamber slides上培养的黏附细胞冰冻或福尔马林固定、石蜡包埋样本分析时间:2-3小时,除外培养、固定和渗透检测次数:一个试剂盒50T试剂盒存储/稳定性:未开封试剂盒储存于-15~-25℃可稳定至标签上标明的效期。

1 流程图:2 样品准备2.1 黏附细胞、细胞涂片和细胞离心涂片需准备的其他试剂:Washing buffer:磷酸盐缓冲液(PBS)Blocking buffer封闭溶液:甲醇稀释的3% H2O2Fixation solution固定溶液:PBS配制的4%多聚甲醛,ph 7.4,新鲜配制Permeabilisation solution 渗透液:0.1%Triton1)X-100溶于0.1%柠檬酸钠溶液中,新鲜配制步骤:下表描述了细胞固定、内源性过氧化物酶封闭和细胞渗透过程。

2.2 组织部分2.2.1 福尔马林-包埋组织福尔马林包埋组织的预处理:可按4种不同的方式预处理。

临床检验中的分子诊断技术进展考核试卷

临床检验中的分子诊断技术进展考核试卷
B. Bisulfite sequencing
C. Western blot
D. Southern blot
11. 哪种技术可以用于检测单个细胞中的基因表达?( )
A. RNA-Seq
B. Single-cell PCR
C. qPCR
D. Northern blot
12. 以下哪种技术不是基于DNA芯片的检测方法?( )
A. Microarray
B. SNP array
C. CGH array
D. Western blot
13. 在分子诊断中,以下哪种技术用于检测基因重排?( )
A. PCR
B. FISH
C. Southern blot
D. Northern blot
14. 哪种技术可用于检测病原微生物的快速诊断?( )
D. PCR
8. 哪种技术被称为下一代测序技术?( )
A. Sanger sequencing
B. NGS
C. AFLP
D. RFLP
9. 以下哪个不是NGS技术的优点?( )
A. 高通量
B. 成本低
C. 准确性高
D. 速度快
10. 在分子诊断中,以下哪种技术用于检测DNA甲基化?( )
A. PCR
C. Western blot
D. Northern blot
19. 哪种技术可以用于检测miRNA的表达?( )
A. PCR
B. Northern blot
C. RNA-Seq
D. Western blot
20. 在分子诊断中,以下哪种技术用于检测病毒载量?( )
A. PCR
B. ELISA
C. Western blot

认知神经科学常用技术及原理_北京师范大学中国大学mooc课后章节答案期末考试题库2023年

认知神经科学常用技术及原理_北京师范大学中国大学mooc课后章节答案期末考试题库2023年

认知神经科学常用技术及原理_北京师范大学中国大学mooc课后章节答案期末考试题库2023年1.是正常人脑电波的一个基本的特征,与注意、警觉以及很多认知过程相关。

相比于睁眼,闭眼显著增加。

答案:alpha波2.在单细胞转录组测序技术中,获得单细胞的方法之一——显微操作挑选法的特点是:答案:能够看见并准确控制单个细胞的吸取与释放,但通量低,对操作人员的技术要求高3.以下关于ERP的说法错误的是:答案:叠加平均之前的波看起来更加平滑4.第一个发现大脑信号有振荡特性的,是。

答案:Hans Berger5.以下关于TMS说法正确的是:答案:单脉冲TMS最主要的功能用于神经功能诊断6.弥散加权图像的磁敏感系数b值越大,信号衰减量,即弥散加权图像的灰度值。

答案:越多,越低7.关于脑磁图测量设备,下列说法错误的是:答案:SQUID与OPM相比更有生态效应8.EEG和都是直接反映神经的电信号,可以在时间上达到神经电活动的毫秒级别。

答案:MEG9.在弥散张量成像中,基于所估计出来的弥散张量模型,科学家们提出了一些弥散参数指标,其中AD值是:答案:反映轴向弥散程度的轴向弥散系数10.以下关于神经振荡,说法正确的是:答案:振荡与睡眠的不同阶段有很强的相关11.以下哪个不是对BOLD-fMRI进行预处理分析的目的:答案:检测脑功能活动的局部差异12.经颅磁刺激在大脑中产生的电流主要于大脑皮层表面,神经元细胞膜与感应电流方向时,磁场刺激作用更明显。

答案:平行,垂直13.脑电帽主要有:答案:另外3项全部正确14.应用单细胞转录组可以进行哪方面的工作:答案:另外3项全部正确15.在进行fNIRS数据分析时,个体水平分析的主要目的是:答案:从fNIRS数据中计算出个体对不同类型刺激的血液动力学响应指标。

Presage ST2 Assay IFU FDA rev 3. 30pg

Presage ST2 Assay IFU FDA rev 3. 30pg
Presage® ST2 Assay - Instructions for Use ____________________________________________________________
Name and Ordering Information Presage® ST2 Assay Kit REF# BC-1065 Presage® ST2 Controls Kit REF# BC-1066 Indications for Use The Critical Diagnostics Presage® ST2 Assay kit is an in vitro diagnostic device that quantitatively measures ST2 in serum or plasma by enzyme-linked immunosorbant assay (ELISA) in a microtiter plate format. The Presage® ST2 Assay is indicated to be used in conjunction with clinical evaluation as an aid in assessing the prognosis of patients diagnosed with chronic heart failure. The Presage® ST2 Controls, Level 1 and Level 2, are designed to be used for monitoring the performance of test procedures on the Critical Diagnostics Presage® ST2 Assay kit. Summary and Explanation Heart failure is a chronic, progressive disease in which the ability of the heart to provide needed cardiac output weakens, thus impeding the heart’s ability to pump enough blood to support the body’s metabolic demands. The prevalence of heart failure is growing worldwide and is a major burden on hospital care costs. (AHA, 2011). A major component to this burden is the fact that patients afflicted with advanced heart failure have high rates of hospitalization and resource utilization, and similarly have a high risk for death (Schappert et al. 2008). Clinicians continue to seek improved methods to determine prognosis, and risk stratification in heart failure remain inadequate. One method now available to clinicians for such assessment is biomarker testing. ST2 (growth STimulation expressed gene 2; also known as IL1RL1, or Interleukin-1 ReceptorLike 1) is a member of the interleukin-1 receptor family. The ST2 protein has two isoforms directly implicated in the progression of cardiac disease: soluble ST2 and a cell membranebound isoform, ST2L. When soluble ST2 levels are low, ST2’s ligand, IL-33, is available to bind to ST2L and has a cardioprotective effect resulting in preserved cardiac function. However, when soluble ST2 levels are high, soluble ST2 competitively binds to IL-33, making IL-33 less likely to bind to ST2L and thereby making IL-33 unavailable for cardioprotective signaling. As a result, the heart is subjected to greater stress in the presence of high levels of soluble ST2, leading to cellular death and tissue fibrosis, reduced cardiac function, and an increase in the rate of disease progression (Kakkar & Lee 2008; Seki et al. 2009). The Presage® ST2 Assay quantitatively measures the concentration of soluble ST2, providing a physician with an accurate tool to assess prognosis in patients with chronic heart failure.

PacBio SEQUELII 准备 HiFi SMRTbell 库说明书

PacBio SEQUELII 准备 HiFi SMRTbell 库说明书

Procedure & Checklist – Preparing HiFi SMRTbell®Libraries using the SMRTbell Express Template Prep Kit 2.0This procedure describes the construction of HiFi SMRTbell libraries for de novo assembly and variant detection applications using the SMRTbell Express Template Prep Kit 2.0 and recommended HiFi sequencing conditions using PacBio’s new Sequel® II Binding Kit 2.2. A minimum input amount of 5 µg of high-molecular weight genomic DNA is recommended for generating HiFi library yields sufficient for running multiple SMRT®Cells on the Sequel II or Sequel IIe System (Sequel II Systems). Note that final HiFi library construction yields will be dependent on the specific size-selection method employed.We recommend fragmenting the gDNA so that the target size distribution mode is between 15 kb - 18 kb. To reduce the presence of fragments >30 kb, PacBio recommends a 2-cycle shearing method on the Megaruptor 3 system. Generally, a narrower fragment size distribution results in more uniform and higher-quality HiFi data. Details regarding DNA shearing conditions (e.g., buffers and DNA sample concentration) are described in the “DNA Requirements for Shearing” section.RequiredEquipment Vendor Throughput Run TimeFemto Pulse AgilentTechnologies Process up to 11 samples per runBatch process up to 88 samples 85 minsMegaruptor 3 Diagenode Shear up to 8 samples at a time40 mins(for 1 cycle of shearing)PippinHT Sage Science Maximum of 20 samples per instrument run 2 hrsBluePippin Sage Science Maximum of 4 samples per instrument run 4.5 hrsSageELF Sage Science Maximum of 2 samples per instrument run 4.5 hrs Table 1: Recommended equipment for HiFi SMRTbell library construction for de novo assembly and variant detection applications.Required MaterialsDNA SizingFemto Pulse Agilent Technologies, Inc. P-0003-0817DNA QuantitationQubit™ Fluorometer ThermoFisher Scientific Q33238Qubit 1X dsDNA HS Assay Kit ThermoFisher Scientific Q33230DNA ShearingMegaruptor 3 System Diagenode B06010003Megaruptor 3 Shearing Kit Diagenode E07010003SMRTbell Library PreparationSMRTbell® Express Template Prep Kit 2.0 PacBio 100-938-900AMPure® PB Beads PacBio 100-265-900SMRTbell® Enzyme Clean Up Kit 2.0 (New*) PacBio 101-932-600Sequencing Primer v5 (New*) PacBio 102-067-400100% Ethanol, Molecular Biology Grade Any MLSWide Orifice Tips (Tips LTS W-O 200UL Fltr RT-L200WFLR) Rainin 30389241Lo-Bind 0.2 mL tube strips USA Scientific, TempAssure1402-4708Multi-channel Pipette Rainin, 17013810Magnetic separation rack V&P Scientific, Inc, VP 772F4-1Thermal Cycler that is 100 µL and 8-tube strip compatible Any MLSSize-selection (One of the following systems)PippinHT System Sage Science HTP00010.75% Agarose Gel Cassettes, Marker 75E Sage Science HPE7510BluePippin System Sage Science BLU00010.75% Agarose Cassettes, Marker S1 Sage Sciences BLF7510SageELF System Sage Science ELF00010.75% Agarose Cassettes Sage Science ELD7510SequencingSequel® II Binding Kit 2.2 (New*)PacBio 101-894-200Sequel® II Sequencing Kit 2.0 PacBio 101-820-200SMRT® Cell 8M Tray PacBio 101-389-001* To obtain a copy of the previous version of this Procedure & Checklist that specifies use of SMRTbell Enzyme Clean Up Kit (PN 101-746-400) and Sequencing Primer v2 (PN 101-847-900), contact ****************.HiFi Library Construction WorkflowPacBio recommends that gDNA samples be resuspended in an appropriate buffer (e.g., Qiagen Elution Buffer) before proceeding with DNA shearing.Figure 1: Workflow for preparing HiFi libraries using the SMRTbell Express Template Prep Kit 2.0.Reagent HandlingSeveral reagents in the SMRTbell Express Template Prep Kit 2.0 (shown in Table 2 below) are sensitive to temperature and vortexing. We recommend to:•Never leave reagents at room temperature.•Always work on ice when preparing master mixes.•Finger-tap followed by a quick spin prior to use.Reagent Where UsedDNA Prep Additive Remove single-strand overhangsDNA Prep Enzyme Remove single-strand overhangs DNA Damage Repair Mix v2 DNA Damage RepairEnd Prep Mix End-Repair/A-tailingOverhang Adapter v3 LigationLigation Mix LigationLigation Additive LigationLigation Enhancer LigationSMRTbell Enzyme Clean Up Mix Nuclease TreatmentSMRTbell Enzyme Cleanup Buffer 2.0 Nuclease TreatmentTable 2: Temperature sensitive reagentsGenomic DNA (gDNA) Quality EvaluationThis procedure requires high-quality, high-molecular weight input gDNA with a majority of the DNA fragments >50 kb as determined by pulsed-field gel or capillary electrophoresis. Any of the three commercially available systems listed in Table 4 below may be used to evaluate gDNA quality, but the Femto Pulse system is highly recommended for high-throughput library construction due to its ability to rapidly process multiple samples in a single run using very low amounts (<1 ng) of DNA per sample. Links to recommended procedures for each system are also provided in the table. Examples of gDNA quality assessment using Bio-Rad’s CHEF Mapper (2A) and Agilent Technologies’ Femto Pulse (2B) are shown in Figure 2. Lanes A3 and B1 correspond to high-quality gDNA samples that are suitable for HiFi library construction using this procedure. Lanes A4 and B2 show degraded gDNA samples that not suitable for use in this procedure.Method ProcedureFemto Pulse Agilent Technologies, Inc.Bio-Rad CHEF Mapper XA Pulsed Field Electrophoresis System Procedure & Checklist - Using the BIO-RAD® CHEF Mapper® XA Pulsed Field Electrophoresis SystemSage Science Pippin Pulse Procedure & Checklist - Using the Sage Science PippinPulse Electrophoresis Power Supply SystemTable 3. gDNA Quality Evaluation Methods and Procedures.Figure 2: Evaluation of high-molecular weight gDNA quality using two DNA sizing analysis systems. A) Bio-Rad CHEF Mapper and B) Agilent Technologies’ Femto Pulse.165.510 kb 50 kb 42 kb33 kb 21 kb 17.7 kb 1.3 kb1 bpLane 1: 8 kb - 48 kb Ladder (Bio-Rad) Lane 2: 5 kb ladder (Bio-Rad) Lane 3: HMW gDNA Lane 4: Degraded gDNALane 1: HMW gDNALane 2: Degraded gDNA Lane 3: 165 kb ladder48 kb-20 kb-80 kb----------10 kb-14322 1 3ABDNA Requirements for ShearingBefore shearing, ensure that the genomic DNA is in an appropriate buffer (e.g.,Qiagen Elution Buffer, 10 mM Tris-Cl, pH 8.5 or PacBio EB buffer). If you are unsure of the buffer composition or if the gDNA is not in Elution Buffer, perform a 1X AMPure PB bead purification followed by elution with Elution Buffer or an equivalent low salt buffer (i.e., 10 mM Tris-Cl, pH 8.5- 9.0).PacBio highly recommends Diagenode’s Megaruptor 3 system for shearing gDNA. The Megaruptor 3 system allows up to 8 gDNA samples to be processed simultaneously with a consistent fragment size distribution across multiple hydropore-syringes. Furthermore, the Megaruptor 3 system generates a narrower size distribution than the g-TUBE device (Covaris).Shearing Using Diagenode’s Megaruptor 3 SystemTo maximize HiFi yield per SMRT Cell, PacBio recommends fragmenting the gDNA to a size distribution mode between 15 kb – 18 kb for human whole genome sequencing. Libraries with a size distribution mode larger than 20 kb are not recommended for HiFi sequencing. Recommended library insert size distributions to use for different WGS applications are summarized in Table 4 below.Application Recommended Library Insert SizeHuman Variant Detection 15 – 18 kbHuman de Novo15 – 18 kbPlant/Animal de Novo15 – 20 kbTable 4: Library size recommendations for Human variant detection and de novo assembly.To shear gDNA on the Megaruptor 3 system, use a two-cycle shear method, which requires running a second round of shearing immediately following the first fragmentation step in the same hydropore-syringe. The recommended concentration is 83.3 ng/µL (5 µg of input DNA in 60 µL Elution Buffer).The DNA shearing guidelines below have been tested by PacBio on the Megaruptor 3 system only. The response of individual gDNA samples to the shearing recommendations described below may differ; therefore, performing a small-scale test shear is highly recommended, including the Megaruptor 3 system.For the Megaruptor and Megaruptor 2 systems, shearing optimization is necessary before proceeding with this Procedure & Checklist. The shearing procedure described in the “Shearing Using Diagenode’s Megaruptor 3 system” section below is not compatible with the Megaruptor or Megaruptor 2 systems. For Megaruptor and Megaruptor 2 systems, follow Diagenode’s DNA shearing recommendations described in their manual. For additional guidance, contact Technical Support or your local FAS.The g-TUBE device generates a broader DNA fragment size-distribution compared to the Megaruptor 3 system. Note that HiFi read quality and overall HiFi data yield may be reduced due to the residual presence of large DNA fragments generated by g-TUBEs. For additional guidance, contact Technical Support or your local FAS.Figure 3: Examples of human genomic DNA samples sheared to a target 15 kb - 18 kb size distribution mode using a 2-cycle shear method on the Megaruptor 3 system.Prepare SMRTbell LibrariesAlways work on ice throughout the library construction process. To process multiple samples at a time, the following equipment are required:• Lo-Bind tube strips• Multi-channel pipette• Wide-bore tips• Magnetic rack compatible with tube strips• Thermocycler compatible with tube stripsRemove Single-Strand OverhangsThe sample volume recovered from the Megaruptor 3 system after shearing is used directly in the single-strand overhang digestion step. Before proceeding, ensure that the sheared DNA is in Elution Buffer or an equivalent low salt buffer (i.e., 10 mM Tris-Cl, pH 8.5- 9.0). In this step, DNA Prep Additive is diluted first followed by digestion. Scale up the reaction volumes for digestion if working with multiple samples.1. Prepare the DNA Prep Additive. The DNA Prep Additive is diluted with Enzyme Dilution Buffer toa total volume of 5 µL. This amount is sufficient for processing 1 to 4 samples. The volume maynot be sufficient for 5 samples due to pipetting errors. We recommend scaling up the dilutionvolume based on the number of samples to be processed (example: prepare 2X volume for 8samples and 4X volume for 16 samples).Note: The diluted DNA Prep Additive should be used immediately and should not be stored.2. Prepare the digestion by following the reaction table below. For multiple samples, prepare amaster mix, followed by addition of 10.0μL master mix to each sheared DNA sample.3. Add 10.0 µL of the above master mix to the tube-strips containing 45.0 µL - 53.0 µL of shearedDNA. The total volume in this step is 55.0 µL - 63.0 µL.4. Using a multi-channel pipette, mix the reaction wells by pipetting up and down 10 times with wide-orifice pipette tips.5. Spin down the contents of the tube strips with a quick spin in a microfuge.6. Incubate at 37°C for 15 minutes, then return the reaction to 4°C.7. Proceed to the next step.Repair DNA DamageTo each Reaction Mix 1, add 2.0 µL of DNA Damage Repair Mix v2.1. Mix the reaction well by pipetting up and down 10 times with wide-orifice pipette tips.2. Spin down the contents of the tube strips with a quick spin in a microfuge.3. Incubate at 37°C for 30 minutes, then return the reaction to 4°C.4. Proceed to the next step.End-Repair/A-tailingTo each Reaction Mix 2, add 3.0 µL of End Prep Mix.1. Mix the reaction well by pipetting up and down 10 times with wide-orifice pipette tips.2. Spin down the contents of the tube strips with a quick spin in a microfuge.3. Incubate at 20°C for 10 minutes.4. Incubate at 65°C for 30 minutes, then return the reaction to 4°C.5. Proceed to the next step.Adapter LigationIn this step, 5.0 µL of Overhang Adapter is added to each Reaction Mix 3 (from the previous step). Then, 32.0 µL of the ligase master mix is added to each Reaction Mix 3/Adapter Mix for incubation. Always work on ice. 1. To each Reaction Mix 3, add 5.0 µL of Overhang Adapter.2. Mix the reaction well by pipetting up and down 10 times with wide-orifice pipette tips. Leave the tube strips on ice.3. Prepare a Master Mix containing Ligation Enhancer, Ligation Additive and Ligation Mix using the table4. Mix the reaction well by pipetting up and down 10 times with wide-orifice pipette tips. It is important to mixwell.5. To the Reaction Mix 3/Adapter Mix, add 32.0 µL of the Ligase Master Mix. The total volume in this step is97.0 µL- 105.0 µL.6. Mix the reaction well by pipetting up and down 10 times with wide-orifice pipette tips. It is important to mixwell.7. Incubate at 20°C for 1 hour. Optional: The Ligation reaction may also be left at 20°C overnight.8. Proceed to the next step.Purify SMRTbell Library Using 1.0X AMPure® PB BeadsPage 11 PN 101-853-100 Version 05(August 2021)Nuclease Treatment of SMRTbell LibraryTo each library sample, add the nuclease mix to remove damaged SMRTbell templates.1. Prepare a Master Mix of the Enzyme Cleanup Mix and Buffer.2. Mix the reaction well by pipetting up and down 10 times with wide-orifice pipette tips. It is important to mixwell.3. Spin down the contents of the tube strips with a quick spin in a microfuge.4. To each 15.0μL of sample, add 55.0 μL of Nuclease Master Mix. The total reaction volume at this step is70.0 µL.5. Mix the reaction well by pipetting up and down 10 times with wide-orifice pipette tips. It is important to mixwell.6. Incubate at 37°C for 30 mins and store on ice immediately.7. Spin down the contents of tube strips with a quick spin in a microfuge.8. Proceed directly to the AMPure PB bead purification step below immediately. Do not store samples at thisstage. Do not let samples sit for long periods of time. Always work on ice.Page 12 PN 101-853-100 Version 05(August 2021)Purify SMRTbell Library Using 1.0X AMPure® PB BeadsSize Selection of SMRTbell LibrariesFor high-throughput whole genome sequencing applications, PacBio highly recommends the PippinHT system (Sage Science) for size-selection of SMRTbell libraries for HiFi sequencing. Typical recovery yields are 35% - 50% and are highly dependent on the size distribution of the starting SMRTbell library.Size Selection Using the PippinHT SystemVerify that your PippinHT system software is up to date and follow the procedure below to remove SMRTbellSize Selection Using the BluePippin SystemSage Science’s BluePippin system may also be used for size-selection of HiFi SMRTbell libraries. Verify that your BluePippin system software is up to date and follow the procedure below to remove SMRTbell templates <10 kb using the BluePippin system. Typical recovery yields are highly dependent on the size distribution of the starting SMRTbell library. For the latest BluePippin system User Manual and guidance on size-selection protocols, contact Sage Science ().Size Selection Using the SageELF SystemSage Science’s SageELF system may also be used to fractionate SMRTbell libraries for HiFi whole genome sequencing applications. Verify that your SageELF system software is up to date and follow the size selection procedure below. For the latest SageELF User Manual and guidance on size-selection protocols, contact Sage Science ().6Set up the run Protocol:– In the “Protocol Editor” tab, click on the “New Protocol” button.– Select the “0.75% 1-18kb v2” in the cassette definition menu.– Select “size-based” for separation mode.– Enter 3450 in the “Target Value” field and move the bar slider to selectwell #12.– Save as new protocol.– On the Main screen, clear previous run data, select cassette description,cassette definition and protocol, enter sample ID(s).– Select in the Nest Selector the cartridge that will be run.7Start the run.8 Once the run is complete, (approximately 4.5 hours), collect 30 μL of the respectivefractions from the elution wells. Fractions of interest are typically ~11 kb, ~13 kb,~15 kb, ~17 kb.9 Check the sizes of all 12 fractions by loading on a Femto Pulse. To determine theaverage library size, perform a smear analysis by selecting the region of interestby defining the start and end points of the fractions.10 Pool together fractions that have an average library size 10 – 20 kb.11 Proceed to the AMPure PB Bead purification step.Purify Size-Selected HiFi Library Fractions with 1.0X AMPure ® PB BeadsSequencing PreparationSee Quick Reference Card - Loading and Pre-Extension Recommendations for Sequel II/IIe Systems .For Research Use Only. Not for use in diagnostic procedures. © Copyright 2020 - 2021, Pacific Biosciences of California, Inc. All rights reserved. Information in thisdocument is subject to change without notice. Pacific Biosciences assumes no responsibility for any errors or omissions in this document. Certain notices, terms, conditions and/o r use restrictions may pertain to your use of Pacific Biosciences products and/or third p arty products. Please refer to the applicable PacificBiosciences Terms and Conditions of S a le and to the applicable license terms at /lice nses.html. Pacific Biosciences, the Pacific Biosciences logo, PacBio, S M RT, SMRTbell, Iso-Seq and Sequel are trademarks of Pacific Biosciences. Femto Pulse and Fragment Analyzer are trademarks of Agilent Technologies. All other trademarks are the sole property of their respective owners.Revision History (Description)Version Date Initial release.01 September 2019 Internal revision with no content change (not uploaded to website).02 December 2019 On page 1, changed “HiFi reads” to just “Reads”. On page 12, under Repair DNA Damage,corrected “remove single strand overhangs” to “repair DNA damage”. On page 13, corrected “remove single strand overhangs” to “adapter ligation”.03 January 2020 Updated for SMRTbell Enzyme Clean Up Kit 2.0 and Sequencing Primer v5.04 April 2021 Removed SMRT Link Sample Setup and Run Design tables. Added reference to QRC.05August 2021。

断裂标记原位测序(bliss) 原理 简书

断裂标记原位测序(bliss) 原理 简书

断裂标记原位测序(bliss) 原理简书全文共四篇示例,供读者参考第一篇示例:断裂标记原位测序(BLISS)是一种新兴的高通量测序技术,可以在染色体水平上高效地检测DNA双链断裂位点。

这种技术的原理基于DNA双链断裂位点与同一染色体上的位置发生联结,通过对联结进行测序,可以确定DNA断裂发生的位置以及频率,从而实现全基因组的高通量测序。

BLISS技术的原理主要包括以下几个步骤:样品中的DNA双链断裂位点被修复,并与同一染色体上的相邻位置形成连接。

随后,通过核酸测序技术,可以将这些连接点进行测序,确定断裂点的位置和频率。

通过对测序数据进行分析,可以得到DNA双链断裂在整个基因组上的分布情况,从而揭示DNA的损伤和修复过程。

通过BLISS技术,研究人员可以实现对整个基因组的高通量测序,揭示DNA损伤和修复的动态过程。

该技术在研究DNA损伤与修复的机制、基因组稳定性、肿瘤发生等领域具有广泛的应用前景。

BLISS技术也可以为精准医学和研究人员提供更多的工具和数据支持,促进疾病的诊断和治疗。

第二篇示例:断裂标记原位测序(BLISS)是一种用于研究染色体二级结构和基因组空间组织的革命性技术。

相比传统的染色体捕获测序技术(如3C、4C、5C等),BLISS技术具有更高的灵敏度和分辨率,能够准确、高效地检测染色体上不同位置之间的连接情况,揭示基因组内部的空间结构和相互作用关系。

BLISS技术的原理主要包括以下几个步骤:断裂标记、适配体连接、PCR扩增和高通量测序等。

BLISS技术通过在细胞核内引入DNA断裂酶(如DNAse I)来切断染色体上的DNA链。

这些断裂位点可以反映出染色体上的不同区域之间的连接情况,从而揭示基因组的空间结构。

接着,将合适的适配体连接到DNA链的断裂位点上,使得每个位点都带有一个唯一的标记序列,以便后续的PCR扩增和测序分析。

然后,通过PCR扩增技术扩增含有断裂标记的DNA片段,使得这些片段可以被高通量测序仪准确地测序。

白细胞介素检测

白细胞介素检测

迪信泰检测平台
白细胞介素检测
白细胞介素(Interleukin),又称介白素,是一组细胞因子(分泌的信号分子),可以由多种细胞产生。

最早发现在白细胞中表达作为细胞间信号传递的手段。

免疫系统的功能在很大程度上依赖于白细胞介素。

一些罕见的白细胞介素缺陷不足都常导致自体免疫性疾病或免疫缺陷。

迪信泰检测平台基于高稳定性、可重复和高灵敏度的分离、表征、鉴定和定量分析系统,结合LC-MS/MS提供可靠、快速且经济高效的白细胞介素分析服务。

此外,我们还提供其他类花生酸分析的检测服务,以满足您的不同需求。

LC-MS/MS测定白细胞介素样本要求:
1. 请确保样本量大于0.2g或者0.2mL。

周期:2~3周。

项目结束后迪信泰检测平台将会提供详细中英文双语技术报告,报告包括:
1. 实验步骤(中英文)。

2. 相关参数(中英文)。

3.质谱图片。

4. 原始数据。

5. 白细胞介素含量信息。

迪信泰检测平台可根据需求定制其他物质测定方案,具体可免费咨询技术支持。

罗弗公司2010年版低功耗高电压负压电子传输器2SB1182和2SB1240产品说明书

罗弗公司2010年版低功耗高电压负压电子传输器2SB1182和2SB1240产品说明书

Medium power transistor (-32V, -2A)2SB1182 / 2SB1240●Features1) Low V CE(sat).V CE(sat) = -0.5V (Typ.) (I C /I B = -2A / -0.2A)2) Complements 2SD1758 / 2SD1862.●StructureEpitaxial planar type PNP silicon transistor●Absolute maximum ratings (Ta=25︒C)∗1 Single pulse, Pw =100ms∗2 Printed circuit board, 1.7mm thick, collector copper plating 100mm 2or larger.ParameterV CBO V CEO V EBO P C Tj Tstg−40V V V A(DC)W (Tc =25°C )W°C °C−32−5−2I CA (Pulse)−3101∗1∗22SB11822SB1240150−55 to 150Symbol Limits Unit Collector-base voltage Collector-emitter voltage Emitter-base voltage Collector currentCollector power dissipationJunction temperature Storage temperature●Electrical characteristics (Ta=25︒C)∗ Measured using pulse current.ParameterSymbol BV CBO BV CEO BV EBO I CBO I EBO h FE V CE(sat)f T CobMin.−40−32−5−−120−−−−−−−−10050−−−−1−1390−0.8∗∗−−V I C = −50μA I C = −1mA I E = −50μA V CB = −20V V EB = −4V I C /I B = −2A/ −0.2A V CE = −5V, I E =0.5A, f =100MHz V CB = −10V, I E =0A, f =1MHzV VμA μA −−V CE = −3V, I C = −0.5AV MHz pFTyp.Max.Unit Conditions−0.5Collector-base breakdown voltage Collector-emitter breakdown voltage Emitter-base breakdown voltage Collector cutoff current Emitter cutoff currentCollector-emitter saturation voltage DC current transfer ratio Transition frequency Output capacitance●Packaging specifications and h FEPackage CodeBasic ordering unit (pieces)TapingTL 2500h FE QR 2SB1182−TV22500−QR2SB1240Typeh FE values are classified as follows :Item h FEQ 120 to 270R 180 to 390●Electrical characteristic curvesFig.1 Grounded emitter propagationcharacteristicsBASE TO EMITTER VOLTAGE : V BE (V)C O L L E C T O R C U R R E N T : I C (m A )−−−−−−−−−−Fig.2 Grounded emitter outputcharacteristics−−−−−C O L LE C T O R C U R R E N T : I C (A )COLLECTOR TO EMITTER VOLTAGE : V CE (V)Fig.3 DC current gain vs.collector curren ( )D C C U R RE N T G A I N : hF ECOLLECTOR CURRENT : I C (mA)Fig.4 DC current gain vs.collector current ( )D C C U R RE N T G A I N : hF ECOLLECTOR CURRENT : I C (mA)Fig.5 Collector-emitter saturationvoltage vs. collector current ( )C O L L E C T O R S A T U R A T I O N V O L T A G E : V C E (s a t ) (m V )−−−−Fig.6 Collector-emitter saturationvoltage vs. collector current ( )−−−−−C O L L E C T O R S A T U R A T I O N V O L T A G E : V C E (s a t ) (m V )COLLECTOR CURRENT : I C (mA)Fig.7 Base-emitter saturation voltagevs. collector currentCOLLETOR CURRENT : I C (mA)B A S E S A T U R A T I O N V O L T A G E : V B E (s a t )(V )−−−−−Fig.8 Gain bandwidth product vs.emitter currentEMITTER CURRENT : I E (mA)T R A N S I T I O N F R E Q U E N C Y : f T (M H z )Fig.9 Collector output capacitance vs.collector-base voltageEmitter input capacitance vs. emitter-base voltageCOLLECTOR TO BASE VOLTAGE : V CB (V)EMITTER TO BASE VOLTAGE : V EB (V)C O L L E C T O R O U T P U T C A P AC I T A N C E : C o b (p F )E M I T T E R I N P U T C A P A C I T A N C E : C i b (p F )Fig.10 Safe operation area(2SB1182)−−−−−−−−−−C O L L E C T O R C U R R E N T : I C (A )COLLECTOR TO EMITTER VOLTAGE : V CE (V)NoticeN o t e sNo copying or reprod uction of this d ocument, in part or in whole, is permitted without theconsent of ROHM Co.,Ltd.The content specified herein is subject to change for improvement without notice.The content specified herein is for the purpose of introd ucing ROHM's prod ucts (hereinafter"Products"). If you wish to use any such Product, please be sure to refer to the specifications,which can be obtained from ROHM upon request.Examples of application circuits, circuit constants and any other information contained hereinillustrate the standard usage and operations of the Products. The peripheral conditions mustbe taken into account when designing circuits for mass production.Great care was taken in ensuring the accuracy of the information specified in this document.However, should you incur any d amage arising from any inaccuracy or misprint of suchinformation, ROHM shall bear no responsibility for such damage.The technical information specified herein is intended only to show the typical functions of andexamples of application circuits for the Prod ucts. ROHM d oes not grant you, explicitly orimplicitly, any license to use or exercise intellectual property or other rights held by ROHM andother parties. ROHM shall bear no responsibility whatsoever for any dispute arising from theuse of such technical information.The Products specified in this document are intended to be used with general-use electronicequipment or devices (such as audio visual equipment, office-automation equipment, commu-nication devices, electronic appliances and amusement devices).The Products specified in this document are not designed to be radiation tolerant.While ROHM always makes efforts to enhance the quality and reliability of its Prod ucts, aProduct may fail or malfunction for a variety of reasons.Please be sure to implement in your equipment using the Products safety measures to guardagainst the possibility of physical injury, fire or any other damage caused in the event of thefailure of any Product, such as derating, redundancy, fire control and fail-safe designs. ROHMshall bear no responsibility whatsoever for your use of any Product outside of the prescribedscope or not in accordance with the instruction manual.The Prod ucts are not d esigned or manufactured to be used with any equipment, d evice orsystem which requires an extremely high level of reliability the failure or malfunction of whichmay result in a direct threat to human life or create a risk of human injury (such as a medicalinstrument, transportation equipment, aerospace machinery, nuclear-reactor controller, fuel-controller or other safety device). ROHM shall bear no responsibility in any way for use of anyof the Prod ucts for the above special purposes. If a Prod uct is intend ed to be used for anysuch special purpose, please contact a ROHM sales representative before purchasing.If you intend to export or ship overseas any Product or technology specified herein that maybe controlled under the Foreign Exchange and the Foreign Trade Law, you will be required toobtain a license or permit under the Law.Thank you for your accessing to ROHM product informations.More detail product informations and catalogs are available, please contact us.ROHM Customer Support System/contact/。

碧云天 细胞凋亡-一步法TUNEL检测试剂盒

碧云天 细胞凋亡-一步法TUNEL检测试剂盒

一步法TUNEL细胞凋亡检测试剂盒产品简介:碧云天生产的一步法TUNEL细胞凋亡检测试剂盒(One Step TUNEL Apoptosis Assay Kit)为您提供了一种高灵敏度又快速简便的细胞凋亡检测方法。

对于经过固定和洗涤的细胞或组织,只要经过一步染色反应,洗涤后就可以通过荧光显微镜或流式细胞仪检测到呈现绿色荧光的凋亡细胞。

细胞在发生凋亡时,会激活一些DNA内切酶,这些内切酶会切断核小体间的基因组DNA。

细胞凋亡时抽提DNA进行电泳检测,可以发现180-200bp的DNA ladder。

基因组DNA断裂时,暴露的3’-OH可以在末端脱氧核苷酸转移酶(Terminal Deoxynucleotidyl Transferase, TdT)的催化下加上绿色荧光探针荧光素(FITC)标记的dUTP(fluorescein-dUTP),从而可以通过荧光显微镜或流式细胞仪进行检测,这就是TUNEL(T dT-mediated d U TP N ick-E nd L abeling)法检测细胞凋亡的原理。

注:FITC是fluorescein isothiocyanate的缩写,实际上大多数情况下所谓的FITC即为fluorescein。

本试剂盒有如下优点。

(1) 高灵敏度:可以在单细胞水平检测到细胞凋亡,同时由于凋亡早期就有DNA断裂,可以检测到早期的细胞凋亡。

(2) 特异性:TUNEL检测时通常更容易标记凋亡细胞,而不容易标记坏死细胞。

(3) 快速:仅需约1-2个小时即可完成。

(4) 方便:只需一步染色反应,洗涤后即可观察,不必使用二抗等进行多步操作。

(5) 应用范围广:可以用于检测冷冻或石蜡切片中的细胞凋亡情况,也可以检测培养的贴壁细胞或悬浮细胞的凋亡情况。

TUNEL法特异性检测细胞凋亡时产生的DNA断裂,但不会检测出射线等诱导的DNA断裂(和细胞凋亡时的断裂方式不同)。

这样一方面可以把凋亡和坏死区分开,另一方面也不会把射线等诱导发生DNA断裂的非凋亡细胞判断为凋亡细胞。

Synopsys STAR Memory System 测试、修复和诊断解决方案说明书

Synopsys STAR Memory System 测试、修复和诊断解决方案说明书

SYNOPSYS IP DATASHEET/ip The Synopsys Self-Test and Repair (STAR) Memory System™ is a comprehensive, integrated test, repair and diagnostics solution that supports repairable or nonrepairable embedded memories across any foundry, process node or memory IP vendor. Silicon-proven in over a billion chips on a range of process nodes, the STAR Memory System is a cost-effective solution for improving test quality and repair of manufacturing faults found in advanced technologies like FinFET. The STAR Memory System’s highly automated design implementation and diagnostic flow enables SoC designers to achieve quick design closure and significantly improve time-to-market and time-to-yield in volume production. The STAR Memory System has been certified for the ISO 26262 automotive functional safety standard by SGS-TUV Saar GmbH, an independent accredited assessor. In addition, the test and repair support for e-flash and embedded MRAM (eMRAM) enables the STAR Memory System to be used in IoT applications.SoC designers, silicon aggregators, and leading foundries targeting automotive, IoT, enterprise, and consumer applications license STAR Memory System with the added flexibility of consulting services for memory BIST planning, generation, insertion, and verification.STAR Memory System Yield Accelertor Figure 1: Synopsys STAR Memory System SolutionHighlights• Complete memory test, repair anddiagnostics solution supportingembedded SRAM, register files, CPU andGPU caches, CAM, multi ports, embeddedflash, MRAM as well as external memorysuch as DDR/LPDDR• Increased design productivity withhierarchical architecture and automatedsystem-on-chip (SoC) integration andverification• High-quality test to provide full memorydefect coverage with minimum test time• High yield with efficient on-chip repairacross multiple operating corners• Superior diagnostics with physical failedbitmaps and XY coordinate identificationto quickly determine root cause of failures• Increased field reliability with STARMemory System Compiler for ErrorCorrecting Codes’ (ECC) multi-bit transienterror correction• ISO 26262 certified to meet the safetyrequirements of high reliability designstargeting up to and including the moststringent ASIL D standard• Supports Internet of Things (IoT)applications with the industry’s firstcommercial built-in self-test (BIST)solution for embedded flash andembedded MRAM (eMRAM)STAR Memory System SolutionFigure 2: The Synopsys STAR Memory System helps save millions of dollars in recovered silicon,reduces test costs, and shortens time-to-volumeThe STAR Memory System Solution Consists of:• Synthesizable test and repair register transfer level (RTL) IP• STAR Memory System Design Acceleration (DA) scripts: Automates the planning, generation, insertion, and verification of test and repair RTL IP• STAR Memory System Yield Accelerator: Automates the generation of tester ready patterns in WGL/STIL/SVF, test algorithm programmability and post silicon failure diagnostics and fault classification• STAR Memory System Silicon Browser: Provides interactive silicon debug of memory using a personal computer or workstation • STAR Memory System ECC: Automatically generates ECC Verilog code, testbenches and scripts for single-port and multiport SRAM memories• STAR Memory System ext-RAM: Offers a high-coverage, cost-effective test with optional Post Package Repair (PPR) and diagnostics solution for external memories such as DDR, LPDDR and HBM via JTAG, either during production or in-field test • STAR Memory System CAM: Supports specialty content addressable memory (CAM) such as binary, ternary and XYCAMs with support for common CAM capabilities• STAR Memory System eMRAM: Optimizes manufacturing yield for eMRAM technology with inclusion of NVM specific test algorithms and support for trimming in addition to test, repair and diagnosisFlexible, Open SystemTo provide STAR Memory System access to all memory developers, Synopsys offers a specialized memory description language called MASIS. The MASIS language, together with a MASIS compiler, simplifies and automates the process of creating and verifying memory views used by the STAR Memory System. By providing an open interface to the STAR Memory System, Synopsys extends the value of the system to all users regardless of whether or not they elect to use Synopsys memories (Figure 2).High-Performance Core SupportThe STAR Memory System allows at-speed test and repair of high-performance processor cores by using a preconfigured test bus, which provides access to the memories inside the core in test mode. The STAR Memory System uses this shared Multi-Memory Bus (MMB) to test memories and add memory test and repair logic outside the IP core to avoid any impact on processor core performance (Figure 1).Test Algorithm ProgrammabilityThe STAR Memory System provides full test algorithm programmability. The STAR Memory System processor includes a BIST module to execute test algorithms. The default test algorithms in the BIST module can be replaced with new algorithms in the RTL or in silicon, and the user can program either their own custom algorithms or select from the comprehensive library of algorithms provided in the STAR Memory System.2Tester Patterns and DiagnosticsThe STAR Memory System Yield Accelerator addresses the need to identify, analyze, isolate and classify memory faults as designs are readied for transition from first silicon to volume manufacturing rapidly, cost-effectively and accurately. Leveraging the infrastructure of the STAR Memory System, the Yield Accelerator automatically generates vectors for test equipment and provides fault analysis and root-cause failure guidance based on silicon test results. Using this feature, test and product engineers can rapidly analyze failures manifested in embedded memories and inspect the physical location and class of each fault to determine the root cause without involving the IP vendor or SoC designer.On-Chip Self-RepairUnlike complex external repair flows, the STAR Memory System’s on-chip repair is fully automated. A built-in self-diagnosis module determines the location of any memory defect and provides error logging by scanning out failure data for silicon debug. When testing memories with redundancies that have failures, a built-in repair and redundancy allocation module identifies available redundant elements and determines the best possible redundancy configuration.Integrated Test and Repair with Synopsys Embedded MemoriesBy hardening the timing-critical test and repair logic within the memory hard macro, the Synopsys STAR Memory System provides unique integration with Synopsys Embedded Memory Compilers. Optimal placement of the timing-critical test and repair logic near the memory allows faster design closure, higher performance, better area and reduced power.Silicon Bring-up and CharacterizationThe STAR Memory System Silicon Browser has advanced automation capabilities to interactively communicate through a JTAG port with the STAR Memory System’s infrastructure in a chip for post-silicon bring-up, system debug, diagnosis and characterization of embedded memories. The unique features of the Silicon Browser allow full extraction of memory contents, multi-corner and multi-voltage characterization, precise physical failure localization, defect classification and redundancy utilization analysis, all from an engineer’s desktop, and without the need for expensive automatic test equipment.©2023 Synopsys, Inc. All rights reserved. Synopsys is a trademark of Synopsys, Inc. in the United States and other countries. A list of Synopsys trademarks isavailable at /copyright.html. All other names mentioned herein are trademarks or registered trademarks of their respective owners.02/27/23.CS1057939197-Update-to-STAR-Memory-System-DS.。

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Abstract - We present a conmercial logic BIST diagnostic approach; using an enhanced BIST controller that supports sim-ple synchronised observation of the MISR on a per pattern basis, reinitialising the BIST controller. Selected patterns are unloaded and diagnostic inference identifies candidate faults and their locations. The approach is flexible and can be tailored to fit con-straints of the test environment. At-speed BIST is supported, as is diagnostics through a low speed interface, such as IEEE 1149.1.I IntroductionThis paper presents a pragmatic approach to the extraction of diagnostic information from a failing logic BIST session. Our approach uses an innovative diagnostic mode within the BIST controller to expedite the search for multiple input signature register (MISR) values that observe faults from the design under test (DUT). This removes the need to perform complex searches to identify failing BIST patterns.Logic BIST is mostly used for system test. However, as the hardware is there it can be used as part of the manufacturing test flow too[12]. Being able to locate the source of failures is important during manufacturing test, it can also render useful feedback on reliability issues when system test fails.Logic BIST[1][2] provides a compact pass/fail signature[3] value. Much has been written about extracting diagnostic information from a BIST run[3][4][5][6][7][8][9][10][11]. The weakness of these solutions is their hardware overhead, behaviour in the context of multiple faults in the DUT or the need for multiple BIST runs to extract diagnostic information.A new approach is proposed by Wohl[13], using STIL[14][15] and interval unloads. Our diagnostic approach can be used in a similar manner. However, we are able to pre-screen directly to failing patterns without significant over-head. Also, our BIST controller diagnostic mode provides synchronisation enabling diagnostics to be performed when BIST is being clocked from a free running clock, such as an on chip clock generator (at speed).II BIST Controller Architecture forDiagnosticsOur BIST controller follows the STUMPS[16] architecture and has a state machine that controls the BIST session. The BIST controller is designed for use from an 1149.1[17] com-pliant TAP interface. We provide a two register logical inter-face. A selection register controls the internal register accessed through the data register port. So, to interact with the value of MISR one must load the selection register with MISR_SELECT value and then access the data register. Both the selection register and the data register are fully shadowed and provide synchronisation between the slow clock domain of the logical interface and that of the BIST controller. The exact implementation of this interface is beyond the scope of this paper. Our BIST controller supports scan chain concate-nation, presenting the many short DUT scan chains as a smaller number of longer ones. It can also be configured to present a single concatenated scan chain for TAP access. The BIST controller is designed to run from an at-speed clock, it provides internal clock control and is capable of generating complex at-speed capture clock waveforms and a slow speed shift clock.There are two novel features that provide direct support for the diagnostic process. Our BIST controller can be initialised to run a window over any part of the overall BIST session. When in diagnostic mode the BIST session supports synchro-nisation points, sensitive to activity on the controller’s physi-cal interface.A Restartable BIST ControllerRather than using masking to protect the MISR from being polluted by the unknown scan chain contents sampled during the first pattern of a window we assert a hold control on the MISR.When performing a BIST run over a window we preload the MISR with the predicted initial value for the start of the win-dow and initialise the pseudo random pattern generator (PRPG), the pattern counter and stop value for the window. At the end of a fault free window the MISR will have the expected fault free value. The pattern counter start and stop values are used, rather than just a count of the number of pat-terns as the absolute pattern number controls the selection of capture clock behaviour and test point activation. The MISR, PRPG and DUT scan chain values can be computed once, at BIST insertion time.Logic BIST Diagnostics Using Simple Synchronised MISR UnloadChris Hill and Thomas RinderknechtDesign For Test Group of Mentor Graphics Corporation8005 SW Boeckman Road, Wilsonville, Oregon 97070Fig. 1. BIST and data register synchronisation BDiagnostic Synchronisation PointsWhen the BIST controller is in diagnostic mode it waits, at the end of each BIST pattern, for the logical interface to per-form a data register load/unload shift sequence, as shown in Fig. 1. Once the sequence has been performed the BIST con-troller continues to the next pattern. It is this waiting that pro-vides synchronisation between the execution of the BIST patterns and activity occurring externally that is gathering the information for the diagnostic process.These synchronisation points allow diagnostics to be per-formed from the external interface to the BIST controller when the BIST process is being clocked at-speed from a free running clock source, such as a phase locked loop (PLL) clock generator. All that is required externally is to wait suffi-cient time for the next synchronisation point to have been reached, no other external handshaking is required. This makes it possible to consider diagnostics of an embedded DUT, where only an 1149.1 TAP is available.Using this diagnostic mode we are able to load/unload the MISR and observe it, reloading it with the fault free value.Hence the MISR value observed for each BIST pattern is made independent and no adjustment is required after a fail-ing MISR value has been observed.Another way to use the diagnostic mode is to unload the con-tents of the DUT scan chains, which will contain the values that would be observed in the MISR after the next BIST pat-tern had unloaded them there. So, it is possible for a given range of BIST patterns to fully unload and observe the DUT scan chains. Given sufficient time and storage capabilities a fully exhaustive unload run of the actual BIST patterns could be performed. Note, in this scenario no load data is specified to fill the scan chain contents, as they are fully refilled when the next BIST pattern is triggered. Also, the MISR value is ignoredIII Diagnostic ProcessFig. 2 shows the steps in the process. These steps are described below:1.Run the BIST controller to perform the BIST session At BIST creation time test vectors to run the full BIST session and observe the MISR value are created. These vectors are used. A failure observe the expected MISR value indicates that the DUT contains observable faults.Fig. 2. Four Step Logic BIST Diagnostic ProcessThese full BIST session vectors are compact in nature and perform the operations as shown in the pseudo code below:Reset BIST controllerInitialise BIST controller Start BIST runWait for BIST run to have completedObserve BIST signature and verify it is expected value2.Identify the failing BIST patternsUse the BIST controller in diagnostic mode to identify the BIST patterns that cause the MISR to observe a faulty value. A second set of test vectors, generated at BIST cre-ation time, run the BIST in diagnostic mode. These vec-tors initialise the BIST controller and then perform the actions described in the pseudo code below:Reset BIST controllerInitialise BIST controller For each BIST patternWait for 1 BIST pattern to complete Unload MISR and reload expected value Verify BIST signature is expectedStartBIST Identify failing StopBIST patternsUnload and expect selected failing patternsFault simulation diagnostic inferencesignature pass?noyesProceed with invasiveinvestigationStep 2Step 3Step 4Step 1Run BIST sessionThese vectors are compact, the bulk of them only contain expected MISR values. If this set of vectors is too large for the test environment it can safely be split into a number of subsets. Just like conventional ATPG vector based ATE application these failing pattern discovery vec-tors can be prematurely terminated once a sufficient[21] quantity of failing patterns have been discovered.3.Unload failing pattern contentsThis step requires a third set of vectors, these can be swiftly generated once the failing BIST pattern set has been identified. This step uses the failing patterns, or a subset of them, and a selection of expected passing pat-terns. The vectors of this step use the BIST controller to initialise the scan chains within the core under test and perform the BIST capture cycle. Then, the vectors unload the scan chain contents and comparing them to the expected scan chain contents for the pattern. This is shown in the pseudo code below:Reset BIST controllerFor each failing BIST patternInitialise BIST controller patternWait for BIST pattern to completeUnload core scan chain contentsVerify core scan chain againstexpected valueFor a large number of failing BIST patterns this vector set may become sizable. Since diagnostic inference can pro-duce reliable deductions with a small number of failing patterns, it is expected that only a subset of failing BIST patterns will be unloaded for further analysis.In order to ensure that the BIST controller is operating correctly the vector set contains a number of significant known passing patterns, within the scan chain integrity test1 and each of the MTPI phases. If these patterns are not unloaded correctly then it is most likely that a defect within the BIST hardware is preventing correct operation of the BIST.Access to the DUT scan chains is through either the con-catenated single scan chain or the DUT scan chains pre-sented at the level of the BISTed DUT. When the BIST controller is in diagnostic mode and waiting for synchro-nisation the scan chains are held in scan mode. Clocking the BIST controller’s diag_clk input causes the scan chain to shift once.4.Diagnostic InferenceTake the diagnostic failure map and logic BIST pattern set into fault simulation and run diagnostic inference. This final part of the process is identical to the step that would be performed during conventionall diagnostic analysis using functional or structural vectors, rather than the logic BIST patterns.A Static vs Dynamic Vector GenerationThe process described above requires the generation of test vectors to perform the DUT scan chain unload step, for the BIST patterns that cause the MISR to observe a failing value. This may not be convenient, if for example the BIST pattern, PRPG and MISR database is not available at this time. In such circumstances a more conventional window based search of the BIST pattern space and unload of the DUT scan chain contents for the failing windows can be employed. The vector sets for this operation can be created at BIST creation time. In this case step two and three of the process above would be replaced with:2.Identify the failing BIST pattern regionsDivide the BIST pattern space into a number of regions2.Run the BIST controller to locate the BIST pattern space regions that cause MISR signature to diverge.This second collection of vector sets is static in nature and can be generated during the BIST controller generation process. Each of these vector sets are very compact, con-taining only initialisation, partial run and a MISR signa-ture unload. Their behaviour is as shown in the pseudo code below:Reset BIST controllerInitialise BIST controller for windowRun BIST controller for windowUnload MISR valueVerify expected MISR value for window end The size of the region is determined by the capabilities of the ATE and the limitations placed on the unload described in the next step. If a BIST pattern region is found to fail, producing a mismatch with expected signa-ture when the vectors are run, then it can be added to the collection of windows for further investigation.3.Unload failing pattern regionsUse the BIST controller to initialize the scan chains in the core under test with each BIST pattern in the failing region and unload and compare the scan chains with expected values.The vectors of this step use the BIST controller to initial-ise the scan chains within the core under test and perform the BIST capture cycle. Then, the vectors unload the scan chain contents and comparing them to the expected scan chain contents for the pattern. This is shown in the pseudo code below:Reset BIST controllerFor each failing BIST patternInitialise BIST controller for patternWait for BIST pattern to completeUnload DUT scan chain contentsVerify DUT scan chainThe vectors required for this step are static in nature and can be generated during BIST controller generation proc-1.During these patterns the scan enable input is held high duringthe capture cycle, so that scan shifting problems can easily be identified.2.These BIST pattern regions should be sized to fit the constraintsof the ATE.ess. The previous step identifies exactly which vector sets, from the sets for all regions of the BIST pattern space, need to be exercised on the ATE. Normally sufficient diagnostic information will be contained within the first few failing BIST pattern regions.As previously, in order to ensure that the BIST controller is operating correctly one would like to observe at least some passing patterns unloaded from the failing BIST pat-tern regions. If these patterns are not encountered one can not be certain that the failures are not caused by a defect within the BIST controller.B Using 1149.1 TAPThe BIST controller does not use the 1149.1 RUNBIST instruction. A BIST session is controlled directly from the data registers within the BIST controller. To initiate a BIST session one needs only to load the RUN_BIST value into the BIST controller’s selection register. When this value is in the selection register the MISR is accessible through the data register. The BIST session will run through to completion, or until the next diagnostic synchronisation point, unless the value in the selection register is changed. Once enabled, oper-ation of the BIST is independent of the TAP.When running in diagnostic mode all that is needed to syn-chronise to the BIST controller is a suitable delay, so that by the time the TAP reaches the Capture-DR state, with the BIST data register access instruction loaded, the BIST controller has reached the next synchronisation point. Passing through the Update-DR state triggers the next BIST pattern. If access-ing the DUT scan chain contents, through the single scan chain, one must force a visit to Updated-DR TAP state for BIST controller’s data register. A fake access to the MISR, will accomplish this.IV Advantages of this ApproachA Using BIST vectorsDuring the diagnostic process we use the BIST hardware to create and apply each pattern to the DUT. Observation is then done either using the MISR or the directly through the DUT scan chains. In this way any fault observable to the BIST can be diagnosed and the full capability of the BIST hardware to exercise the faults is available, including at-speed clock con-trol and complex capture windows. No complex test environ-ment is required for the unit under test. No diagnostic resolution is lost when a failing part is located on an embed-ded subsystem with limited access, through 1149.1 TAP or some other test interface.B FlexibilityUsing the restartable nature of the BIST controller one can perform a traditional search of the BIST pattern space, using independent windows, for the failing BIST patterns. Using the diagnostic synchronisation mode of the BIST controller allows easy observation of the MISR value or DUT scan chain contents for a BIST window of BIST patterns.Both of these capabilities put together allow one to tailor one’s diagnostic approach to the exact constraints of the test interface and test environment being used at diagnostic time.A full BIST run can be used with diagnostic synchronisation and MISR unload to determine the BIST patterns that observe failures in the MISR. These failing patterns can then be rerun and the actual DUT scan chain contents observed.If the constraints of the test access method are exceeded by this direct approach then the BIST pattern space can be divided into windows and the approach used within these windows. Kapur[18] suggests using non uniform window sizes, where the window is sized to include the a similar number of new fault detects. Our overall BIST architecture and the use of multi-phased test point insertion (MTPI)[19] would suggest that initial window sizes coincident with the test point phase size should be considered.C Leveraging ATPG Diagnostic InferenceDiagnostic inference, from regular scan based test vectors, is a mature art. Our diagnostic approach leverages on this capa-bility. Once a set of diagnostic unload vectors have been exer-cised using the BIST hardware and the observed failures reported this failure information is mapped into a format that can be taken directly into fault simulator based diagnostic inference, with respect to the original BIST patterns. This process then produces the candidate fault sites for the observed failures.As our approach uses the actual DUT core scan chain values captured by the failing BIST patterns there is no diagnostic resolution lost. The same resolution is available as would be if the BIST patterns and capture procedures had been applied directly to the DUT. Our approach is effective irrespective of the number of faults within the DUT. The entire diagnostic approach is independent of the fault model being used. It works just as well for stuck-at-faults as for transition-faults. Being that we are aiming to make the BIST patterns as diag-nosable as conventional ATPG patterns the difference between fault models is a matter for fault simulation alone.D Vector VolumeBIST runs tend to require a large volume of pattern data, given the pseudo random nature of that data. Compare this to the volume of test data required for a similar fault coverage when using deterministic ATPG vectors. As an example we took a design of 32,000 gates (6,000 scan elements) with 342 inputs and 208 outputs. ATPG was able to reach 98.2% fault coverage in just 383 scan patterns. This represents a total vec-tor volume ofsymbols.To achieve 97.0% fault coverage with BIST required the use of 4,096 BIST patterns. In this example we used a 64 bit38334220860002×++()×4806650=MISR, hence the controller has a 64 bit data register. To run the entire BIST session requires the scan loading of 5 BIST data register values and the final observation of the MISR.This represents symbols. The MISR unload patterns for the entire BIST run would require the same amount of initialisation data then the observation of 4095 MISR values, this represents symbols. Finally, unloading the DUT scan chains for a mixture of 64 failing and passing patterns would requiresymbols. This repre-sents a reduction of the diagnostic data volume by aproxi-mately 90%. Compare this to the requirement of ordinary interval unloads, using a window size of 256 patterns, which would require symbolsto evaluate all of the windows and thensymbols to fully unload each failing window, where each unloaded window will render at least one failing pattern. To obtain the first fail-ing pattern would require about twice the data volume as our approach.E AliasingAliasing of MISR values, a sequence of faulty DUT scan cellvalues causing the MISR to observe a fault free value is a well understood and studied problem in BIST[20]. Given that the MISR is proportioned to have an acceptably low probabil-ity of aliasing over the entire BIST pattern space the probabil-ity of aliasing occurring within the compression of a single BIST pattern is further reduced. Typically a single fault will be observed in multiple failing BIST patterns and hence multiple failing MISR values. The probability of aliasing occurring for each MISR value that would otherwise observe the failure rapidly tends towards zero, because of the conjuction of the respective single pat-tern aliasing probabilities.FFailure MappingPresently there is no standard format for ATE failure informa-tion to be passed off to other tools. Each test environment produces a unique representation of any captured failure information. This problem is currently solved, on an ad hoc basis, by those performing diagnosis using regular functional or structural test vectors. A mapping must be made from the ATE failure information back into the domain of the source test patterns, suitable for consumption by fault simulation for diagnostic inference.With BIST diagnostics this mapping must be made back not to the applied test vectors but to the underlying BIST patterns. Our BIST diagnostic approach currently uses meta-comments and annotations within the source test vectors to maintain the information needed to perform this mapping. When a stand-ard approach for driving this information forward into the test environment is finalised this will be used instead. Failure mapping takes the reported time of a failure, whichcan be either a tester cycle count or absolute time value, and converts this into information more meaningful to the diag-nostic step being performed. During the diagnostic MISR unload steps is just a matter of determining which BIST pat-tern is triggering the failing MISR value to be observed. For the DUT scan chain content unload step the failure mappingmust relate the failure back to the DUT core scan chain and element position within it. This mapping may need to take account of any scan chain concatenations present in the BISTed DUT, even being able to map back from the singlescan chain used through the 1149.1 TAP.V Disadvantages of this ApproachAs we stated above, to make effective use of the diagnostic unloading of MISR values and then to be able to unload onlythe targeted failing patterns and the selection of passing pat-terns, requires access to the BIST database of pattern, MISRand PRPG values. If this is not possible at the point at which the diagnostic data is being gathered then this approach will not work. One can fall back to the alternative window based unload approach also outlined above.Running diagnostics through the 1149.1 tap interface may take some time, considering the volume of data to move.However, extraction of diagnostic information outside of the manufacturing test flow is not a time critical task.Performing BIST diagnostics using the MISR unload mecha-nism outlined above is disruptive, each failing part willrequire it’s own failing pattern unload vector set. Hence, it can only be performed as an off line activity. Using the non disruptive interval unload approach may require a large amount of response/failure storage, because of the data vol-ume involved.The MISR unload mechanism will not work when transient defects 1 cause failures, unless these defects are repeatable using an isolated pattern. Diagnostic information for such defects can be captured using a windowed BIST pattern run terminating on the pattern that observes the defect effect. The DUT scan chains can then be unloaded to locate the scan ele-ment that made the observation. Multiple BIST runs may be necessary to locate the failing pattern.VI ConclusionThe novel hardware within the BIST controller allows the BIST to be run using any number of independent windows of arbitrary size. It also provides a diagnostic mode that allows5644+()×64+409=5644+()×409564×+262420=645644+()×6000+()×405760=4095256-----------5464+()×64+()×6462≈5464+()×2566000×+1536340= 1.Such as power supply noise, triggered by activity in previous pat-terns.access to the MISR and DUT scan chain contents on a per BIST pattern basis, with exceedingly simple synchronisation semantics, without the need to reinitilise the BIST controller for each pattern. These capabilities when used with our sup-porting software tools provide a very flexible diagnostic envi-ronment that can be used to extract complete diagnostic information in a wide range of flows, from a very low data volume binary search and unload of failing patterns through to a high volume unload of all patterns (with or without the use of windowing).We also propose a new technique that allows failing patterns to be identified and then targeted directly for DUT scan chain observation, using the diagnostic mode of the BIST control-ler.The diagnostic approach is valid in situations where there is good access to the failing part as well as those where the fail-ing part is embedded and access is only available through a constrained test interface, such as 1149.1 TAP. Because of the flexibility of the approach the diagnostic process can be tai-lored to match the constraints of the test access method and testing environment.The diagnostic approach provides the same level of diagnos-tic resolution as would be available had the BIST patterns been applied directly to the DUT. It can be used off line on failing parts during manufacturing test or as a reliability diag-nosis tool for parts that fail after manufacture.VII References1. C Stroud, Automated BIST for Sequential Logic Synthesis,IEEE Design and Test of Computers 19882. B Koenemann, T Mucha and G Zwiehoff, Built in LogicBlock Observation Techniques, Proceedings of the Inter-national Test Conference 19793.W H McAnney and J Savir, There is Information in FaultySignatures, Proceedings of the International Test Confer-ence 19874.Y Wu and S Adham, BIST Fault Diagnosis in Scan-BasedVLSI Environments, Proceedings of the International Test Conference 19965.Y Wu and S Adham, Scan Based BIST Fault Diagnosis,IEEE Transactions on Computer-Aided Design of Inte-grated Circuits and Systems, V ol. 18, No 2 February 1999 6.J Rajski and J Tyszer, On the Diagnostic Properties ofLinear Feedback Shift Registers, IEEE Transactions on Computer-Aided Design Oct 19917.J Rajski and J Tyszer, Diagnosis of Scan Cells in BISTEnvironment, IEEE Transactions on Computers V ol 48, No 7 July 19998.J Ghosh-Dastidar, D Das and N Touba, Fault Diagnosis inScan Based BIST Using Both Time and Space Informa-tion, Proceedings of the International Test Conference 19999.J Ghosh-Dastidar and N Touba, A Rapid and ScalableDiagnosis Scheme for BIST Environments with a Large Number of Scan Chains, Proceedings of the VLSI Test Symposium 200010.J Savir, Salvaging Test Windows in BIST Diagnostics,IEEE Transactions on Computers V olume: 47 Issue: 4 April 199811.J Savir, Salvaging Test Windows in BIST Diagnostics, Pro-ceedings of the VLSI Test Symposium 199712.Xijiiang Lin, R Press, J Rajski, P Reuter, T Rinderknecht,B Swanson and N Tamarapalli, High-frequency, at-speedscan testing, Design & Test of Computers, IEEE , V ol-ume: 20 , Issue: 5 , Sept.-Oct. 200313.P Wohl, J Waicukauski, S Patel and G Maston, EffectiveDiagnostics Through Interval Unloads in a BIST Environ-ment, Proceedings of the Design Automation Conference 200214.IEEE Standard Test Interface Language (STIL) for digitaltest vector data, IEEE Std 1450-199915.P Wohl and N Biggs, P1450.1: STIL for the simulationenvironment, Proceedings of the VLSI Test Symposium 200016.P.H. Bardell, W.H. McAnney, and J. Savir, Built-In Testfor VLSI: Pseudo Random Techniques, Wiley Inter-science, New York, 198717.IEEE standard test access port and boundary - scanarchitecture, IEEE Std 1149.1-199018.R Kapur, T Williams and M Mercer, Directed-BinarySearch in Logic BIST Diagnostics, Proceedings of the Design, Automation and Test in Europe Conference and Exhibition 200219.N Tamarapalli and J Rajski, Constructive multi-phase testpoint insertion for scan-based BIST, Proceedings of the International Test Conference 199620.S K Gupta, Recent advances in BIST, VLSI Test Sympo-sium, 1992. '10th Anniversary. Design, Test and Applica-tion: ASICs and Systems-on-a-Chip', Digest of Papers., 1992 IEEE, V ol., Iss., 7-9 Apr 199221.Zhiyuan Wang, Kun-Han Tsai, M Marek-Sadowskal, JRajski, An efficient and effective methodology on the mul-tiple fault diagnosis, Proceedings of the International Test Conference 2003。

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