一种简单有效且适于土壤微生物多样性分析的DNA提取方法  

A Simple and Effective Method of DNA Extraction from Soil for Molecular Biodiversity Analysis

作　　者：张海燕[1] 王彩虹[1] 龚明福[1] 张利莉[1] 

机构地区：[1]塔里木大学生命科学院新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室,阿拉尔843300

出　　处：《生物技术通报》2009年第8期151-155,共5页Biotechnology Bulletin

基　　金：国家"973"计划前期研究专项资助课题"新疆连作棉田土壤微生态平衡机理研究(2007CB116303);新疆生产建设兵团基础研究项目"新疆棉花连作条件下土壤微生物与棉花相互作用机理的研究(2007JC06);新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室开放课题(TD06005)

摘　　要：参照Zhou[11]的方法进行了改进,获得了一种简单、有效的DNA提取方法。此方法操作简单、从大量样品改为小量样品的提取,利用高浓度的PEG沉淀,不作回收纯化,所提DNA片段较大,在23kb以上,每克土的DNA提取量从3.74～15.28μg,OD260/OD230比值在0.89～1.21范围内,用真菌和细菌核糖体特异性引物进行PCR扩增,均获得较好的结果,DGGE图谱显示丰富性较高,可用于细菌多样性和真菌多样性的分析。此方法能够从4种不同性质土壤中提取出DNA,但提取盐渍土壤和碱性土壤的效果更好一些,为土壤微生物群落结构的多样性分析奠定良好的基础。In this paper, based on Zhou＇method, a simple and effective method for direct extraction of DNA was obtained, in which small amount of samples and DNA precipitation with higher concentration polyethylene glycol without purification involved in. The result showed that this method not only generate the largest DNA fragments in size, but also was directly used as PCR amplification with bacterial and fungal specific primers. The OD260/OD230 ratio of the isolated DNA was 0.89 - 1.21, and the yields from one gram soil was 3.74 - 15.28 μg. DGGE profiles demonstrated the method was suit for simultaneous recovery of bacterial and fungal community total DNA. It proved that this method can extracted DNA from four kinds of soil, especially saline-alkaline soil.

关 键 词：土壤微生物多样性 宏基因组DNA DNA提取PCR DGGE 

分 类 号：S154.3[农业科学—土壤学]