基于PCR-DGGE和16S rDNA克隆技术研究土壤细菌多样性的实验技术  

Experiment technology of soil bacterial diversity by using PCR-DGGE and 16S rDNA clone technique

作　　者：刘秉儒[1] 

机构地区：[1]宁夏大学西北土地退化与生态恢复国家重点实验室培育基地,宁夏银川750021

出　　处：《农业科学研究》2014年第3期34-38,共5页Journal of Agricultural Sciences

基　　金：国家自然科学基金项目"贺兰山土壤微生物多样性对植被垂直分异的响应机制"(41061003);宁夏大学博士科研启动基金项目(012-60020342)

摘　　要：PCR-DGGE是测定土壤微生物多样性常用的技术手段之一,但是提供的微生物多样性信息不够完整,如果与16S rDNA克隆技术相结合,微生物多样性信息更加丰富.PCR-DGGE和16S rDNA克隆实验技术复杂,许多新手没有成熟可靠的经验可借鉴.以土壤细菌多样性研究方法为例,分别从DNA提取与纯化、PCR扩增、DGGE、胶片染色技巧以及16S rDNA克隆技术等环节进行技术体系构建,可供利用该技术的研究人员参考.PCR-DGGE has been widely used for measuring soil microbial diversity, but microbial diversity information is limited for identification of microbial communities. If experiment method could completely combined with16 S rDNA clone technique, its limitations could reduce, and conclusions also are accurate, reliable and with good repeatability. But the technology is technical process is more complex, and technical requirements is higher. In order to afford alutary experience or lessons to many new operators, experiment technology from the extraction of DNA, PCR amplification, DGGE experiment technology, dyeing and clone technique were described respectively.

关 键 词：PCR-DGGE 16S RDNA 克隆技术 土壤细菌多样性 实验技术 

分 类 号：S154.3[农业科学—土壤学]