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Mao Dandan, Xu Peiya. Comparison of extraction methods of activated sludge microbial DNA for PCR-DGGE analysis[J]. Chinese Journal of Environmental Engineering, 2013, 7(3): 1189-1195.
Citation: Mao Dandan, Xu Peiya. Comparison of extraction methods of activated sludge microbial DNA for PCR-DGGE analysis[J]. Chinese Journal of Environmental Engineering, 2013, 7(3): 1189-1195.
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Comparison of extraction methods of activated sludge microbial DNA for PCR-DGGE analysis

  • 1.

    College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310014, China

  • Received Date: 09/03/2012
    Accepted Date: 10/11/2011
    Available Online: 18/03/2013
    Fund Project:
  • The aim of this study was to develop a method of extracting total microbial DNA from activated sludge for PCR-DGGE analysis. Five methods were used to extract total microbial DNA from activated sludge. The DNA purity, yield, fragment size, and microbial diversity reflected by DGGE technique were used to evaluate the extracted DNA. The results of UV absorbance and agarose gel electrophoresis showed that the yield of DNA extracted by the kit method was lowest among the five methods. There was no significant difference in the yield of DNA for the other four DNA extraction methods. In terms of the DNA purity, the kit method was the best one. Four DNA extraction methods could effectively lysis both bacterial and fungal cells except the high temperature lysis method for fungal cells. According to the DGGE analysis, more diverse bacterial communities were obtained with the DNA extracted by the high temperature lysis method, and more diverse fungal communities were obtained with the DNA extracted by the SDS lysis method. High temperature lysis method could be more suitable for bacterial PCR-DGGE analysis of activated sludge, while SDS lysis method could be more suitable for fungal PCR-DGGE analysis.
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Comparison of extraction methods of activated sludge microbial DNA for PCR-DGGE analysis

  • Received Date: 09/03/2012
  • Accepted Date: 10/11/2011
  • Available Online: 18/03/2013
Fund Project:

Abstract: The aim of this study was to develop a method of extracting total microbial DNA from activated sludge for PCR-DGGE analysis. Five methods were used to extract total microbial DNA from activated sludge. The DNA purity, yield, fragment size, and microbial diversity reflected by DGGE technique were used to evaluate the extracted DNA. The results of UV absorbance and agarose gel electrophoresis showed that the yield of DNA extracted by the kit method was lowest among the five methods. There was no significant difference in the yield of DNA for the other four DNA extraction methods. In terms of the DNA purity, the kit method was the best one. Four DNA extraction methods could effectively lysis both bacterial and fungal cells except the high temperature lysis method for fungal cells. According to the DGGE analysis, more diverse bacterial communities were obtained with the DNA extracted by the high temperature lysis method, and more diverse fungal communities were obtained with the DNA extracted by the SDS lysis method. High temperature lysis method could be more suitable for bacterial PCR-DGGE analysis of activated sludge, while SDS lysis method could be more suitable for fungal PCR-DGGE analysis.

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