湿地土壤微生物DNA提取及其脱腐技术  

DNA Extraction and Removing Humic Substance from Wetland Soil

作　　者：李靖宇[1] 赵吉[2] 边玉 武琳慧[2] 于景丽[2] 

机构地区：[1]内蒙古大学生命科学学院,内蒙古呼和浩特010021 [2]内蒙古大学环境与资源学院,内蒙古呼和浩特010021

出　　处：《微生物学通报》2010年第8期1130-1137,共8页Microbiology China

基　　金：国家973计划前期研究专项项目(No.2009CB125909)

摘　　要：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。DNA-based molecular biology techniques have widely been used as a powerful tool to understand the microbial community.In this paper,a new method to extract microbial genomic DNA from wetland soil was established,namely Calcium Chloride-SDS-Enzymatic.Calcium chloride rather than EDTA chelating agent was used to remove humic acids in the process of direct DNA extraction.The extracting time is less than 4 hours.In comparing with other two methods,this method is more efficient in removing humic acids from wetland soil,and the purity of extracted DNA is higher which can be applied to PCR amplification.It provides an efficient technology to extract and purify microbial genome DNA from soil for microbial ecological studies.

关 键 词：湿地土壤 DNA提取 腐殖酸 PCR扩增 氯化钙-SDS-酶法 

分 类 号：S154.3[农业科学—土壤学]