Computational analysis of stochastic heterogeneity in PCR amplification efficiency revealed by single molecule barcoding.

ABSTRACT: The polymerase chain reaction (PCR) is one of the most widely used techniques in molecular biology. In combination with High Throughput Sequencing (HTS), PCR is widely used to quantify transcript abundance for RNA-seq, and in the context of analysis of T and B cell receptor repertoires. In this study, we combine DNA barcoding with HTS to quantify PCR output from individual target molecules. We develop computational tools that simulate both the PCR branching process itself, and the subsequent subsampling which typically occurs during HTS sequencing. We explore the influence of different types of heterogeneity on sequencing output, and compare them to experimental results where the efficiency of amplification is measured by barcodes uniquely identifying each molecule of starting template. Ou

SUBMITTER: Best K 

PROVIDER: S-EPMC4602216 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature