PCR ‰©¢‍§ç‰©ç„ˆ†‍

download PCR  ‰©¢‍§ç‰©ç„ˆ†‍

of 60

  • date post

    24-Jan-2016
  • Category

    Documents

  • view

    92
  • download

    3

Embed Size (px)

description

PCR 扩增产物的分析. Gel electrophoresis Restriction endonuclease Molecular hybridization Nucleic acid sequence. PCR 扩增产物的电泳分析. 琼脂糖凝胶电泳 常用于临床检测(定性) 聚丙烯酰胺凝胶电泳 分离效果比琼脂糖好,条带比较集中 可用于科研及检测分析. 琼脂糖凝胶电泳. 琼脂糖凝胶电泳是一种非常简便、快速、最常用的分离纯化和鉴定核酸的方法 琼脂糖是从海藻中提取的一种线状高聚物 根据琼溶解温度,把琼脂糖分为一般琼脂糖和低熔点琼脂糖 - PowerPoint PPT Presentation

Transcript of PCR ‰©¢‍§ç‰©ç„ˆ†‍

  • PCR Gel electrophoresisRestriction endonucleaseMolecular hybridizationNucleic acid sequence

  • PCR

  • 626537DNADNA

  • DNA

    (%)DNA(Kb)0.35600.61200.70.8100.90.571.20.961.50.2312.00.12

  • Tris-TBETris-TAETris-TPETBETPEDNATPEDNADNATAETAE EDTA DNAPCR

  • 10TBETris 108EDTA 9.3 55H2O 1000mlpH8.08.20.5TBE20

  • DNA

    0.6%1%1.4%2%1Kb0.6Kb0.2Kb0.15Kb

  • DNA

    1%1.4%2Kb1.6Kb

  • 400DNA

  • ethidium bromideEBEB0.5ug/mlEB1015minEBDNA>5ng30min

  • (Ag+)Ag+EB200DNA

  • 200400 bp DNA1.21.7%

  • 0.5TBE100ml60(EB)0.5 TBE2mm

  • DNA0.2DNA1 5V/cm 200400bp PCR50V2040min

  • 1KbDNADNADNA0.2%500bp1bp

  • TEMEDNNNNNN'

  • DNA

    %bp**3.510020001004605.080500652608.0604004516012.040200307015.025150156020.0101001245

  • .30%29NN1H2O100ml 4

  • 10%1 10 ml 4-201TBETEMED

  • DNA1%45603%TEMED

  • 100.1TBE

  • DNADNA

  • 1-8V/cm0.5ug/ml1530min>10ngDNA

  • DNA Marker DNA100 bp ladder

  • PCR

  • PCRPCR Southern

  • Southern PCR PCR

  • PCR PCR

  • RDB --29AG-28 AG17 ATAAG TAGE CD26 GAG AAGIVS-I-1 GT IVS-I-5 GC27-28+C41-42-CTTT43 GT 71-72+A+T654 CT

  • PCRPCR

  • PrincipleNOS11014/cm2 N-

  • Principle --CCCCCCCapture Probe:NH2Detection Probe:Biotin

  • Principle

  • PrincipleNH2BiotinAvidin-HRP NOS

  • HBV5HBV DNA PCR 32P SouthernPCRELISA

  • PCR

  • DNA

  • DNADNADNA 0.153.0mol/L NaCl

  • DNADNA NaCl NH42SO4 DNADNA

  • DNAMg2+DNADnaseDNA300bp 200ng624bpDNADNA

  • DNADNA 100 5min 10mmol/L-10mmol/L EDTApH7.0NaClDNA372hPBS-0.05%Tween20 3

  • PCR

  • ADNAAB5AB-

  • HLAPCRROXCOUM

  • PCR-ELISAELISA5PCR5PCR5

  • 1 13U/mgSigmaPBS-Tween140mmol/L NaCl 2.7mmol/L KCl4.3mmol/LNa2HPO41.4mmol/L K2HPO40.05%(w/v)Tween20pH7.21g/ml100lPBS

  • 2 PCR1l501 PBS-Tween30minPBS-Tween6

  • 3 ELISAPCR50l PBS-TweenHRP-FITC30minPBS-Tween61mg TMB1ml50mmol/L -pH4.91030%v/vH2O23l80l TMB25min80l 2mol/L ELISA450nmOD

  • PCR-RFLP

  • PCR-SSCP

  • Sequence

  • 2-3FITCDNA DNAGCN5TyrRPCR ELISAPCR-ELISAPCR PCR