Comparison of Total Microbial DNA Extraction Methods from Solid-culture of Zhenjiang Vinegar

Abstract:

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 μg/g dried culture. The quantity of bacteria and fungi reached to the maximum of 1.73×1013 copies?(gram dried culture)-1 and 6.49×1012 copies?(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.