Ni Zhengfei, Xu Wei, Dou Wenfang, Xu Hongyu, Xu Zhenghong
School of Medicine and Pharmaceutics, Jiangnan University, Wuxi 214122, China.
Wei Sheng Wu Xue Bao. 2010 Jan;50(1):119-25.
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.
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).
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.
The results showed that the most effective method was SDS high salt extraction combined with grinding in liquid nitrogen and lysozyme lysis.
分析传统固态发酵过程中微生物的多样性和演替情况。以镇江香醋为例，我们比较了11种从镇江香醋固态培养物中的微生物提取基因组DNA的方法。
采用紫外分光光度计和实时荧光定量PCR测定DNA的产量和纯度。通过变性梯度凝胶电泳（DGGE）分析固态发酵过程中的细菌和真菌多样性。
总DNA的最高提取产量为93.2±1.5μg/g干培养物。细菌和真菌数量分别达到最大值1.73×10¹³拷贝×（克干培养物）⁻¹和6.49×10¹²拷贝×（克干培养物）⁻¹。DGGE显示的群落模式受DNA提取方法的显著影响。其中，基于SDS的六种DNA提取方法产生的DNA条带数量较多。
结果表明，最有效的方法是SDS高盐提取结合液氮研磨和溶菌酶裂解。
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