摘要
利用三种提取方法从内蒙古荒漠草原土壤样品中提取了土壤微生物DNA,得到的DNA片段长度在23 kb以上,提取效果较好无降解.采用试剂盒直接纯化、凝胶回收试剂盒纯化、玻璃珠DNA回收试剂盒SK1111 DNA纯化这三种方法对粗提液进行纯化,结果显示:凝胶回收试剂盒纯化后DNA纯度最高,试剂盒直接纯化可得到较纯的DNA,而玻璃珠DNA回收试剂盒SK1111 DNA纯化的效果不佳,不宜进行后续的PCR扩增.以纯化的土壤微生物DNA为模板扩增了细菌16SrDNA,扩增产物分子量为1.5kb左右.通过比较研究提出了一种适合于从荒漠草原土壤中进行微生物DNA提取、纯化及PCR扩增的有效方法.
Three extraction methods of microbial DNA from the desert grassland soil in Inner Mongolia were performed and compared.The results showed that DNA fragments were extracted with over 23 kb molecular weight by three methods and the DNA samples had no degradation.The yield ratio and purity of the DNA by three different purification methods were different.Method of Gel-plus-column provided more efficient DNA purity than the others.Method of using purification Kit directly was also good.However,method of using SK1111 was not suitable for later PCR amplification.The 16S rDNA gene fragments of bacteria were amplified.So a simple and efficient method to extract,purify and amplify DNA from the desert grassland soil was presented by comparing investigation.
出处
《内蒙古大学学报（自然科学版）》
CAS
CSCD
北大核心
2008年第4期430-434,共5页
Journal of Inner Mongolia University：Natural Science Edition
基金
国家自然科学基金资助项目(30560030)
留学回国人员科研启动项目(2005)
内蒙古自治区高校科研项目(NJ05099)
