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Updated: Jul 3, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
[Influence of different 16S rDNA target sequence on analysis of microbial diversity in anaerobic ABR reactor]
1State Key Laboratory of Water Resource Utilization and Environmental Pollution Control, School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin 150090, China. weilihit@126.com
Abstract:
In the study on denitrification inhibiting sulfate reducing bacterium, it was discussed for the influence of 16S rDNA target sequence from different primers on DGGE fingerprinting from the anaerobic active sludge diversity in ABR reactor. The sludge DNA in the reactor was isolated, four sets primers 341F/534R, 968F/1401R, 63F/534R, and 341F/926R was used to amplify 16S rDNA, and the resolution of DGGE fingerprinting, community diversity were analyzed. The result indicated that it was beneficial to the DNA extraction from sulfate reducing sludge by PBS washing sludge and sonic oscillation; by analysis of DGGE from different primers, there were obvious differences in community diversity. The target sequences from primers 341F/534R and 968F/1401R were isolated relatively well, that from primers 341F/926R was in common, and that from primers 63F/534R was the worst. The DGGE fingerprinting bands from primers 341F/534R were abundant with the best diversity, while that from 968F/1401R was not as well as the former, that from 341F/926R was in common, and that from 63F/534R was the least with worse diversity than the others. Primers 341F/534R and 968F/1401R are recommended to be used simultaneously to analyze anaerobic active sludge by DGGE.
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