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Updated: Oct 19, 2025

Visualizing Methane-Cycling Microbial Dynamics in Coastal Wetlands
Published on: January 31, 2025
Tagged Highly Degenerate Primer (THDP)-PCR for Community Analysis of Methane- and Ammonia-oxidizing Bacteria Based on
Jian-Gong Wang1, Fei Xia1, Jemaneh Zeleke1
1Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, China.
Abstract:
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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