Physiology 3140A Study Guide - Quiz Guide: Housekeeping Gene, Taqman, Exonuclease
29 Dec 2016
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Dna-dependent dna polymerase synthesizes a complementary copy of the template strand. Primers x2 speci c to the dna segment that needs to be ampli ed: process, denaturation. Dsdna are separated via denaturation = increase temperature. Separation of dna strands, 94 100 : annealing. Primers anneal to each strand : forward primer (5-prime primer) & reverse primer (3-prime primer) Allows for primers to bind dna (variable based on the sequence of the primers) Requires a 3"oh prime end for polymerase to operate. Optimal temperature for the polymerase to function: 70 75 : there are 3 stages of pcr ampli cation, exponential phase. Dif cult to distinguish between sample"s original quantities, not enough ampli cation has occurred linear phase. Samples are best compared with each other as they are being ampli ed. The only phase that is sensitive enough to yield data to determine the relative concentration of mrna over time: plateau phase. Ampli cations are no longer ef cient: visualizing pcr.
