一种可用于PCR扩增的直接提取土壤细菌DNA的方法

本文以澳大利亚桉树林和松树林的土壤为例 ,采用Napp提取液和SDS直接溶解土壤细菌 ,并配合温浴 -玻璃珠震荡、苯酚 -氯仿萃取和异丙醇提取以及纯化DNA等步骤 ,直接从土壤样品中提取了土壤细菌DNA。所得DNA完全适用于酶解和PCR扩增的要求。该方法高效简单 ,费用低 ,在土壤微生物研究中具有重要的应用价值

The Method for Bacterial DNA Extraction Directly from Soils for PCR Amplification

An efficient and simple method to obtain DNA from the environment is needed for analysis of bacterial community diversity in soils.A procedure to rapidly extract and purify DNA,being suitable for efficient amplification by PCR,was developed in this paper.Key features of the extraction and purification were fresh and cold Napp buffer,SDS-assisted lysis,with beat beading with interval hot incubate and acid phenol-chloroform and,finally CsCl and Kac precipitation.The resulting DNA was pure enough to be restricted by enzymes and was amplified by PCR.Amplification of the 1500 bp target,16S rDNA from two kinds of forest soils in Australia,was confirmed with a pair of universal primers.Compared to other commonly used DNA extraction methods,it is considered as a rapid and inexpensive way of extracting bacterial DNA from soils directly.