Determining Polymerase Fidelity in the PCR of Mouse HPRT Exon 3
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thesis / dissertation description
The polymerase chain reaction has made it feasible to analyze sections of DNA for mutation because it allows a low number of sequence copies to be amplified to the level of detection. However, since commercially available DNA polymerases do not provide proof-reading capabilities during the PCR reaction, errors are often produced during synthesis. This could produce serious background problems in the detection of naturally occurring mutations in genomic DNA. The author's laboratory had previously confirmed the high error rate of heat-stable polymerase Replinase during PCR. In this study, the author wants to determine whether manual PCR with a heat sensitive modified T7 polymerase would produce a higher fidelity product that then Replinase used in those studies, and whether this improvement would allow sufficient delineation of bands in DGGE to use PCR to study spontaneous and induced mutation rates.