Barcoded Primers Used in Multiplex Amplicon Pyrosequencing Bias Amplification
Open Access
- 1 November 2011
- journal article
- research article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 77 (21), 7846-7849
- https://doi.org/10.1128/aem.05220-11
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.Keywords
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