Effect of the enzyme and PCR conditions on the quality of high-throughput DNA sequencing results.

ABSTRACT: Library preparation protocols for high-throughput DNA sequencing (HTS) include amplification steps in which errors can build up. In order to have confidence in the sequencing data, it is important to understand the effects of different Taq polymerases and PCR amplification protocols on the DNA molecules sequenced. We compared thirteen enzymes in three different marker systems: simple, single copy nuclear gene and complex multi-gene family. We also tested a modified PCR protocol, which has been suggested to reduce errors associated with amplification steps. We find that enzyme choice has a large impact on the proportion of correct sequences recovered. The most complex marker systems yielded fewer correct reads, and the proportion of correct reads was greatly affected by the enzyme used. Mod

SUBMITTER: Brandariz-Fontes C 

PROVIDER: S-EPMC4306961 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature