Evaluation of the impact of single nucleotide polymorphisms and primer mismatches on quantitative PCR.

ABSTRACT:

Background

Robust designs of PCR-based molecular diagnostic assays rely on the discrimination potential of sequence variants affecting primer-to-template annealing. However, for accurate quantitative PCR (qPCR) assessment of gene expression in populations with gene polymorphisms, the effects of sequence variants within primer binding sites must be minimized. This dichotomy in PCR applications prompted us to design experiments to specifically address the quantitative nature of PCR amplifications with oligonucleotides containing mismatches.

Results

We performed qPCR reactions with several primer-target combinations and calculated ratios of molecules obtained with mismatch oligonucleotides to the average obtained with perfect match primer pairs. Amplifications were performed wi

SUBMITTER: Boyle B 

PROVIDER: S-EPMC2741440 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature