Barcoded Primers Used in Multiplex Amplicon Pyrosequencing Bias Amplification

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2011 Sep 6

PMID 21890669

Citations 216

Authors

David Berry

Karim Ben Mahfoudh

Michael Wagner

Alexander Loy

Affiliations

Soon will be listed here.

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.

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