Barcoded primers used in multiplex amplicon pyrosequencing bias amplification.

ABSTRACT: "Barcode-tagged" PCR primers used for multiplex amplicon sequencing generate a thus-far-overlooked amplification bias that produces variable terminal restriction fragment length polymorphism (T-RFLP) and pyrosequencing data from the same environmental DNA template. We propose a simple two-step PCR approach that increases reproducibility and consistently recovers higher genetic diversity in pyrosequencing libraries.

SUBMITTER: Berry D 

PROVIDER: S-EPMC3209180 | biostudies-literature | 2011 Nov

REPOSITORIES: biostudies-literature