聚合酶链式反应 Polymerase Chain Reaction ( PCR)
of 13
description
聚合酶链式反应 Polymerase Chain Reaction ( PCR). 实验三. 实验原理. - PowerPoint PPT Presentation
Transcript of 聚合酶链式反应 Polymerase Chain Reaction ( PCR)
-
Polymerase Chain Reaction (PCR)
-
PCRDNAPCR----DNADNA93DNAPCRDNADNA()DNA55DNADNA--TaqDNAdNTPDNA ----2423(Plateau)
-
PCRPCRDNA16-30bpG+C40-60%Tm4(G+C)+2(A+T)33GC3334 5 51-2
-
4(dNTP)dNTPNaOHpH7.0dNTP5-10mmol/L-20dNTP20-200mol/L4dNTP20mol/L, 100L2.6gDNAdNTP50mmol/LTaq DNA4dNTPdNTP Mg2+Mg2+Taq DNA, Mg2+0.5-2mmol/LPCR0.1-5mmol/LMg2+, Mg2+PCREDTAMg2+Mg2+
-
PCRDNADNADNAPCRDNAPCRTaq DNATaq DNA92.5 130 min95 40min97 5minTaq DNA7430min10nmol/L dNTPTaq DNAPCR210-4 /PCR100L PCR1.5-2Taq DNA30DNA10-50mmol/L TrisCl (20pH8.3-8.8)50mmol/L KClMg2+TrisCl20pH8.3-8.8PCR, pH6.8-7.850mmol/LKCl5mmol/L(DDT)100g/mL(BSA)T432DNATaq DNA
-
PCR
-
PCR50L35L H2O5L 10PCR4L 25mmol/L MgCl2 4L 4dNTP 0.5L 0.5L 0.5L DNA (1ng)
-
PCR94 3min94 30sec50 30sec72 80sec Cycle 4072 10min
-
PCR1.PCR2. PCRPCR3. 4. PCR /100LTE 10000rpm15s, , ::, 300L95, -2030min 10000rpm10min,1mL70,.27mL ddH2O
-
1 2 3 4 5 MMDNA marker123-5DNA
-
EB,EBEBEBDNADNADNA(300-360nmDNAtip
-
,
