湿地土壤微生物DNA提取及脱腐技术.pdfVIP

微生物学通报 AUG 20, 2010, 37(8): 1130−1137 Microbiology China © 2010 by Institute of Microbiology, CAS tongbao@ 研究报告 湿地土壤微生物 DNA 提取及其脱腐技术 1 2* 2 2 2 李靖宇 赵吉 边玉 武琳慧 于景丽 (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) 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4 h 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。 关键词: 湿地土壤, DNA 提取, 腐殖酸, PCR 扩增, 氯化钙-SDS-酶法 DNA Extraction and Removing Humic Substance from Wetland Soil LI Jing-Yu1 ZHAO Ji2* BIAN Yu2 WU Lin-Hui2 YU Jing-Li2 (1. College of Life Sciences, Inner Mongolia University, Huhhot, Inner Monglia 010021, China) (2. College of Environment Resources, Inner Mongolia University, Huhhot, Inner Monglia 010021, China) Abstract: DNA-based molecular biology techniques have widely been used as a powerful tool to un- derstand 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 t