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[Comparison of total microbial DNA extraction methods from solid-culture of Zhenjiang vinegar]
Zhengfei Ni1, Wei Xu, Wenfang Dou
1School of Medicine and Pharmaceutics, Jiangnan University, Wuxi 214122, China. nzf984629@163.com
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.
