Barcoded primers used in multiplex amplicon pyrosequencing bias amplification.
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ABSTRACT:
SUBMITTER: Berry D
PROVIDER: S-EPMC3209180 | biostudies-other | 2011 Nov
REPOSITORIES: biostudies-other
Publications
Barcoded primers used in multiplex amplicon pyrosequencing bias amplification.
Berry David D Ben Mahfoudh Karim K Wagner Michael M Loy Alexander A
Applied and environmental microbiology 20110902 21
"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. ...[more]
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