Barcoded Primers Used in Multiplex Amplicon Pyrosequencing Bias Amplification-Reference-Cited by-同舟云学术

Barcoded Primers Used in Multiplex Amplicon Pyrosequencing Bias Amplification

Published:2011-11 Issue:21 Volume:77 Page:7846-7849
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Berry David¹,Ben Mahfoudh Karim¹,Wagner Michael¹,Loy Alexander¹

Affiliation:

1. Department of Microbial Ecology, Vienna Ecology Center, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria

Abstract

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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/AEM.05220-11

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