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DNA分子生物学技术的广泛应用,为全面了解微生物群落提供了有力的工具。本文建立了一种新的从湿地土壤中提取微生物总DNA的方法,即氯化钙-SDS-酶法。在直接提取DNA过程中采用氯化钙去除腐殖酸,DNA提取缓冲液中不使用EDTA螯合剂,提取过程用时4h左右。与其他两种方法相比,该方法高效去除湿地土壤腐殖酸,纯度较高,满足PCR扩增,为微生物生态学研究提供了一种高效的湿地土壤微生物总DNA提取和纯化技术。
The widespread use of DNA molecular biology techniques provides a powerful tool for understanding microbial communities in all aspects. In this paper, a new method for extracting total DNA from wetland soil was established, that is, calcium chloride-SDS-enzymatic method. In the direct extraction of DNA using calcium chloride to remove humic acid, DNA extraction buffer does not use EDTA chelating agent, the extraction process takes about 4h. Compared with the other two methods, this method can effectively remove humic acid from wetland soils and has high purity and meets PCR amplification. It provides an efficient technique for total DNA extraction and purification of soil microorganisms in microbial ecology.