Barcoded Primers Used in Multiplex Amplicon Pyrosequencing Bias Amplification

被引：402

作者：

Berry, David ^{[1
]}

Ben Mahfoudh, Karim ^{[1
]}

Wagner, Michael ^{[1
]}

Loy, Alexander ^{[1
]}

机构：

[1] Univ Vienna, Dept Microbial Ecol, Vienna Ecol Ctr, Fac Life Sci, A-1090 Vienna, Austria

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2011年 / 77卷 / 21期

基金：

奥地利科学基金会;

关键词：

RARE BIOSPHERE; PCR; DIVERSITY; WRINKLES;

D O I：

10.1128/AEM.05220-11

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

"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.

引用

页码：7846 / 7849

页数：4

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