全自动临床样品核酸的纯化和应用课件

全自动临床样品核酸的纯化和应用生产工具推动社会进步，进步的源泉来源于不断创新。QIAGEN-BioRobot全自动临床样品核酸的纯化和应用生产工具推动社会进步，进步的源QIAGEN公司简介?来自德国，创建于1985年，世界上最优秀的核酸纯化厂商。年销售额数十亿美金，产品畅销全球。?提供各种样品核酸纯化的优质试剂。根据单个样品量不同，有不同的纯化试剂盒；根据样品数量不同，有各种通量的试剂盒。?提供全自动核酸纯化工作站，此工作站同时具备自动移液功能，并可以和其他仪器联合使用，具有不同的扩展功能。QIAGEN公司简介?来自德国，创建于1985年，世界上最优QIAGEN公司仪器简介?BioRobot 9600：通量较低的标准型BioRobot 9604：用于临床样品标准型BioRobot 3000：多功能用户定制型BioRobot 8000：高通量全自动标准型BioRobot RapidPlate：96和384孔板移液系统BioRobot Twister I：存储和移板系统，和 3000连用，也可和酶标仪、PCR仪等连用。?BioRobot Twister II：和8000连用，功能同I型。QIAGEN公司仪器简介?BioRobot 960BioRobot 9604 主要功能?专为临床样品的核酸自动纯化设计，可以得到高纯度的基因组DNA、病毒DNA及RNA。?样品来源：各种血样、体液、口腔刮离细胞及细胞培养物等。?所得到的核酸用于分子诊断、病毒检测，血液分型、法医DNA指纹鉴定、制药公司药物筛选等。BioRobot 9604主要功能?专为临床样品的核酸自动纯BioRobot 9604 主要优点?分子诊断所需DNA/RNA的自动纯化?精确、快速的液面定位系统和移液系统?满意的得率和可信的结果?自动的样品跟踪和过程控制系统?同时进行96样品，且有很好的可重复性?一次性带滤芯枪头可安全处理有传染性的样品?配套的高质量试剂?简单易学的软件操作系统BioRobot 9604主要优点?分子诊断所需DNA/RNBioRobot 9604 亚型?A：Viral RNA?B：Viral RNA/DNA，Genomic DNA，Swab，Amplification Set up。?C：Total RNA，Cytoplamic RNA。BioRobot 9604 亚型?A：Viral RNA?BBioRobot 9604 硬件 介绍14513321261178109BioRobot 9604硬件介绍14513321261171?2?3?4?5X-Y-Z-Robotic Arm：液面定位和自动移液Dilutor Syringe Drives：对移液量的精确检测Vacuum Manifold System：产生负压，从纯化板抽滤掉各种缓冲液Sample identification System：样品条形码读取系统Tip-Disposal Station：去除一次性带滤芯枪头1?2?3?4?5X-Y-Z-Robotic Arm：液面?6Tip-Rack Holder Area：可存放足够的一次性带滤芯枪头?7External Reagent Delivery Module：各种缓冲液存放位置?8High-Speed Pipetting System：控制4个加液头的加液速度?9Labware Identification System：手持型96孔纯化板条形码读码系统?QIAsoft Operation System：简单好学的操10作系统，编好的操作程序，亦可根据需要自己编写程序?6Tip-Rack Holder Area：可存放足够的一Safety Shield：安全防护罩12?Multipurpose Slot：多功能槽，可以放收集管等13?Thermoblock Slot：加热模块，用于升高温度以加快蛋白裂解等?11Safety Shield：安全防护罩12?MultipurQIAsoft Operation System?在WINDOWS 95 基础上编写而成?有编好的现成操作程序，看懂英文就会操作?可以根据自己的需要自行编写程序QIAsoft Operation System?在WINDQIAsoft 3.0 运行环境?Execute Environment：运行已编写好的程序，并有配套的试剂盒提供。?Edit Environment：创建和编写自己的程序。?Setup Environment：用于增加和调整 slot和 rack的位置，与Edit Environment 同时使用。QIAsoft 3.0运行环境?Execute EnviroQIAsoft 3.0 Main Menu?EXCUTE:Start Sample Processing?EDIT:Create Your Own Protocols?SETUP:Adjust Work Table ConfigurationQIAsoft 3.0 Main Menu?EXCUTE:SExecute EnvironmentPROCESS PANELTIME PANELMASSAGE LOGPROCOTOLExecute EnvironmentPROCESS PANBioRobot 9604 样品来源BioRobot 9604 样品来源BioRobot 9604 配套试剂?试剂盒中含有用于结合核酸的硅胶膜板和收集板、封口膜等所有附件。?试剂盒中有足够的各种缓冲液用于纯化所规定的样品数。?每一个试剂盒都有对应的已经编好的程序。BioRobot 9604 配套试剂?试剂盒中含有用于结合核核酸纯化原理核酸（水相）吸附高盐乙醇低PH低盐高温高PH洗脱核酸（结合硅胶膜）核酸纯化原理核酸（水相）吸附高盐乙醇低PH低盐高温高PH洗脱Nucleic Acid in SolutionNucleic Acid in SolutionEffect of Chaotropic SaltsEffect of Chaotropic SaltsBinding of Nucleic AcidBinding of Nucleic AcidQIAamp 8 DNA BLOOD BioRobot Kit(72)QIAamp 96 DNA BLOOD BioRobot Kit(12)?从新鲜或冻存的血液、血清、骨髓、体液、细胞悬液自动纯化基因组、线粒体和病毒的DNA。?2个小时内从200UL样品中得到6UG DNA。?用于一般的分子生物学操作外，还可以用于HLA分型、基因疾病的诊断等。QIAamp 8 DNA BLOOD BioRobot KiQIAamp 96 Virus BioRobot Kit(12)QIAamp 96 Virus RNA BioRobot Kit(12)?从血浆、血清、尿液、脑脊/髓液、无细胞的体液、细胞培养上清液中自动纯化病毒的RNA/DNA。?前者可以在2.5个小时内从200UL样品中同时得到90%以上的DNA和RNA，后者可以在20-40分钟内从140UL样品中得到90%以上的病毒RNA。?所得到的DNA/RNA主要用于病毒检测和分型。如肝炎病毒A-G，HIV，CMV和肠道病毒等。QIAamp 96 Virus BioRobot Kit(1QIAamp 96 DNA Swab BioRobot Kit(12)?从棉签（如口拭子）上纯化DNA?在2个小时内可得到1-2UG DNA?所得到的DNA除可以用于一般的分子生物学操作外，还可以用于 HLA分型、基因疾病的诊断、亲子鉴定等。QIAamp 96 DNA Swab BioRobot KiRNeasy 96 BioRobot Kit(24)?从人和动物细胞中纯化总 RNA和细胞质RNA?可以在 75分钟内从 10-5X106细胞中得到100UG RNA?所得到的 RNA除可以用于一般的分子生物学操作外，还可以用于高通量药物筛选、药物研究和分子诊断等。RNeasy 96 BioRobot Kit(24)?从人自动纯化的过程和应用?因样品不同而大同小异，主要是样品的预处理和所用的缓冲液有所不同，这在各个产品的操作手册中都有详细说明。?以从全血中纯化核酸为例作一些介绍,从200UL样品中得到6UG DNA，纯度可以达到90%以上，并在2个小时内处理完96个样品。自动纯化的过程和应用?因样品不同而大同小异，主要是样品的预处QIAamp 96 DNA BLOOD BioRobot Kit(12)?Number of Preparations 12x96QIAamp 96 plates 12Square-Well Blocks 14Tape Pad 1x25 sheetsAirPore Tape 1x25 sheetsCaps for Collection Microtubes 8x120Collection Microtube Racks 122-ml Tubes for QIAGEN Protease 50Caps for QIAGEN Protease 50QIAamp 96 DNA BLOOD BioRobot KQIAamp 96 DNA BLOOD BioRobot Kit(12)?15-ml Tubes for Buffer AL 15QIAGEN Protease 6x140mgProtease Solvent(sodium azide)6x10ml500-ml Bottle for Buffer AW2 1Buffer AL(chaotropic salt)1x330mlBuffer AW1(concentrate)6x175mlBuffer AW2(concentrate)4x274mlBuffer AE 48x5mlHandbook 1QIAamp 96 DNA BLOOD BioRobot K三种缓冲液的作用?QIAGEN Protease：裂解蛋白。?Protease Solvent(sodium azide)：含叠氮化钠的蛋白溶解液，防止蛋白酶被菌污染。?Buffer AL(chaotropic salt)：在乙醇存在条件下，形成高盐体系，同时蛋白酶在 AL中能更好地裂解细胞，使DNA结合在硅胶膜上。?Buffer AW1(chaotropic salt 和ethanol)：去除硅胶膜上的抑制剂（如血红素、糖酸、肝素、铁离子等PCR抑制剂）。?Buffer AW1（ethanol）：将硅胶膜上的盐去掉（AL、AW1）三种缓冲液的作用?QIAGEN Protease：裂解蛋白。全自动临床样品核酸的纯化和应用课件纯 化 过 程纯化过程水和全血PCR 产物的比较水和全血PCR产物的比较PCR 产物的分析?从水中得到的产物经过PCR后没有扩增出任何产物。?从全血得到的产物经过PCR后扩增出产物，且条带一致，说明重复性很好。PCR产物的分析?从水中得到的产物经过PCR后没有扩增出任何重复性和纯度检验?RNA：A260/A280=1.9-2.1；DNA：A260/A280=1.7-1.9?得率基本上在6ug左右，A260/280=1.8左右重复性和纯度检验?RNA：A260/A280=1.9-2.1HLA Typing?SSP-PCR analysis of donor DNA using primers specific for HLA genes DRB(lanes 123)and DQB(lanes 24 45).Genomic DNA was isolated from human whole blood using the QIAamp 96 DNA Blood BioRobot procedure and amplified with 45 different allele-specific primers targeted at genes DRB1 through to DRB5,and DQB1.Allele-specific amplicons appear as intense bands below the 439 bp control amplicon(human growth hormone gene)in lanes 8,9,22,37,38,and 40.HLA Typing?SSP-PCR analysis ofHuman Leucocyte Antigen TypingHuman Leucocyte Antigen TypingFactor V Analysis?Factor V Leiden genotyping using LightCycle?technology.The figure shows two peaks corresponding to the DNA melting temperatures recorded for the wild-type allele(640C)and the mutant allele(570C)in a heterozygotic individual(red line)and a control DNA(blue line).Genomic DNA was isolated from human whole blood using the QIAamp 96 DNA Blood BioRobot procedure.Factor V Analysis?Factor V LeiFactor V 遗传分析Factor V 遗传分析?END!?THANK YOU VERY MUCH?END!?THANK YOU VERY MUCHI6F3B0y)v%s#pXlUiQfNcK8H5E2A+x*u$rZnWThPeMbJ7G4C1 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