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Comparison among amoA primers suited for quantification and diversity analyses of ammonia-oxidizing bacteria in soil.


ABSTRACT: Ammonia monooxygenase subunit A gene (amoA) is frequently used as a functional gene marker for diversity analysis of ammonia-oxidizing bacteria (AOB). To select a suitable amoA primer for real-time PCR and PCR-denaturing gradient gel electrophoresis (DGGE), three reverse primers (degenerate primer amoA-2R; non-degenerate primers amoA-2R-GG and amoA-2IR) were examined. No significant differences were observed among the three primers in terms of quantitative values of amoA from environmental samples using real-time PCR. We found that PCR-DGGE analysis with the amoA-2IR primer gave the best results in this studied soil. These results indicate that amoA-2IR is a suitable primer for community analysis of AOB in the environment.

SUBMITTER: Shimomura Y 

PROVIDER: S-EPMC4036032 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Comparison among amoA primers suited for quantification and diversity analyses of ammonia-oxidizing bacteria in soil.

Shimomura Yumi Y   Morimoto Sho S   Takada Hoshino Yuko Y   Uchida Yoshitaka Y   Akiyama Hiroko H   Hayatsu Masahito M  

Microbes and environments 20111110 1


Ammonia monooxygenase subunit A gene (amoA) is frequently used as a functional gene marker for diversity analysis of ammonia-oxidizing bacteria (AOB). To select a suitable amoA primer for real-time PCR and PCR-denaturing gradient gel electrophoresis (DGGE), three reverse primers (degenerate primer amoA-2R; non-degenerate primers amoA-2R-GG and amoA-2IR) were examined. No significant differences were observed among the three primers in terms of quantitative values of amoA from environmental sampl  ...[more]

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