[Comparison of total microbial DNA extraction methods from solid-culture of Zhenjiang vinegar]

Wei Sheng Wu Xue Bao. 2010 Jan;50(1):119-25.
[Article in Chinese]

Abstract

Objective: To analyze the diversity and succession of microbes during traditional solid-state fermentation. Taking Zhenjiang vinegar for example, we compared 11 methods for extracting genomic DNA from microbes in solid-culture of Zhenjiang vinegar.

Methods: The yield and purity of the DNA were measured by ultraviolet spectrophotometer and real-time PCR. The bacterial and fungal diversity during solid-state fermentation was analyzed by denaturing gradient gel electrophoresis (DGGE).

Results: The highest extraction yield of total DNA was 93.2 +/- 1.5 microg/g dried culture. The quantity of bacteria and fungi reached to the maximum of 1.73 x 10(13) copies x (gram dried culture)(-1) and 6.49 x 10(12) copies x (gram dried culture)(-1), respectively. The community patterns revealed by DGGE were noticeably influenced by the DNA extraction methods. Hereinto, six methods of SDS-based DNA extraction resulted in the larger number of DNA bands.

Conclusion: The results showed that the most effective method was SDS high salt extraction combined with grinding in liquid nitrogen and lysozyme lysis.

Publication types

  • Comparative Study
  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetic Acid*
  • DNA, Bacterial / isolation & purification*
  • DNA, Fungal / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Fermentation
  • Polymerase Chain Reaction

Substances

  • DNA, Bacterial
  • DNA, Fungal
  • Acetic Acid