Denaturing Gradient Gel Electrophoresis Polymorphism for Rapid 16S rDNA Clone Screening and Microbial Diversity Study

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概要

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• A method based on denaturing gradient get electrophoresis (DGGE) banding polymorphism was developed for rapid small-subunit (SSU) rDNA clone screening and microbial diversity estimation. Clones with correct rDNA insertion were initially identified using whole-cell PCR with vector-specific primers. Subsequently, the correct PCR products were re-amplified with nested primers in a DGGE analysis. An average of five to 10 individual DGGE-PCR products could be mixed together and loaded into a DGGE well, and at least 125 to 250 clones could be screened and compared within one DGGE gel. This approach eliminated the reproducibility problem associated with DGGE, increased the ability to screen a large set of clones at a relatively affordable cost, and was shown to be more sensitive than the restriction fragment length polymorphism method in identifying 10 different but phylogenetically closely related 16S rDNA fragments from 126 clones obtained from an anaerobic sludge sample.

• 公益社団法人日本生物工学会の論文
• 2002-01-25

著者

• Hu Jiang

  Water & Biotreatment Group Department Of Civil Engineering National University Of Singapore

• Liu Wen-tso

  Graduate Institute Of Environmental Engineering National Central University

• Liu Wen-tso

  Graduate Institute Of Environmental Engineering National Central University:water & Biotreatment

• HUANG CHUN-LIN

  Graduate Institute of Environmental Engineering, National Central University

• SONG LIANGFA

  Water & Biotreatment Group, Department of Civil Engineering, National University of Singapore

• ONG SAY

  Water & Biotreatment Group, Department of Civil Engineering, National University of Singapore

• NG WUN

  Water & Biotreatment Group, Department of Civil Engineering, National University of Singapore

• Ng Wun

  Water & Biotreatment Group Department Of Civil Engineering National University Of Singapore

• Ong Say

  Water & Biotreatment Group Department Of Civil Engineering National University Of Singapore

• Song Liangfa

  Water & Biotreatment Group Department Of Civil Engineering National University Of Singapore

• Huang Chun-lin

  Graduate Institute Of Environmental Engineering National Central University

関連論文

• Characterization of a 4-Methylbenzoate-Degrading Methanogenic Consortium as Determined by Small-Subunit rDNA Sequence Analysis
• Isolation, Characterization and Identification of Polyhydroxyalkanoate-Accumulating Bacteria from Activated Sludge
• Denaturing Gradient Gel Electrophoresis Polymorphism for Rapid 16S rDNA Clone Screening and Microbial Diversity Study

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