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Overview
Authored
(1)
JW
Jian-Gong Wang
Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, China
Research fields
Microbiology
Peer-reviewed
Preprint
Tagged Highly Degenerate Primer (THDP)-PCR for Community Analysis of Methane- and Ammonia-oxidizing Bacteria Based on Copper-containing Membrane-bound Monooxygenases (CuMMO)
Authors:
Jian-Gong Wang
,
Fei Xia
,
Jemaneh Zeleke
,
Bin Zou
and
Zhe-Xue Quan
,
date:
06/20/2017,
view:
7828,
Q&A:
0
We describe a two-step PCR strategy using tagged highly degenerate primer (THDP-PCR) targeting copper-containing membrane-bound monooxygenases (CuMMO) genes for community analysis of methane- or ammonia-oxidizing bacteria. This strategy consists of a primary CuMMO gene-specific PCR followed by a secondary PCR with a tag as a single primer. This strategy remarkably increases the divergence of CuMMO gene amplicons while maintaining PCR efficiency without obvious amplification bias. This THDP-PCR strategy can be extended to other functional gene-based community analysis with design of new highly degenerate primer covering target functional gene sequences.
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