重组DNA与遗传工程.pptVIP

EnzymePolymerasebindsinglestrandedDNAatapointadjacenttoadouble-strandedstretch,andbindstodNTPs(deoxynucleotidetriphosphates)ComponentsofPCRreaction:Enzyme(Taq,Vent,Pfu)AppropriatebuffersAmpledNTPsTemplateDNAPrimersCofactors(eg.Magnesium)Thecycle1Denaturation 94or92℃,30sec/incubation Halfactivityafter30incubationsat94℃for60seceach. 60-120secforthefirststepThecycle2Annealing Tm=4(GCbases)+2(ATbases) 30-60sec/incubationThecycle3Extension 72℃,500bp/30sec;500-1500/60secCycles Example: Denature Annealing Ex