摘要
本试验采集蒙古马新鲜粪便样品,采用酚-氯仿抽提法和2种细菌基因组DNA提取试剂盒法进行样品中细菌基因组DNA的提取。通过DNA浓度和纯度的测定,并将其作为模板扩增细菌16SrDNA V3区特异性片段对提取效果进行比较,从而找出更适合蒙古马粪便中细菌基因组DNA的提取方法。结果显示,3种提取方法得到的基因组DNA均能用于PCR扩增的模板,其中B试剂盒法提取的DNA数量较多,且纯度高,更适合用于提取蒙古马肠道微生物细菌基因组DNA及后续的蒙古马肠道微生物多样性等分子生物学试验。
In this study,we extracted total DNA from fresh fecal collected from Mongolian horses through phenol-chloroform technique and two bacterial genomic DNA extraction Kits. To find more suitable methods,we had compared the concentration and purity of DNA and the landscape of amplifying the 16S rDNA V3 region. The results showed that all three methods can be used for DNA extraction from gut bacterial in Mongolian horse. Meanwhile,the quality and purity of DNA extracted by B Kit was the most effective which could be more suitable for the extraction of ge- nomic DNA from gut bacterial and the subsequent analysis of the diversity of bacterial population and biology experiments in Mongolian horse.
出处
《中国畜牧兽医》
CAS
北大核心
2015年第2期498-502,共5页
China Animal Husbandry & Veterinary Medicine
基金
国家科学技术部对俄科技合作专项(2011DFR30860)
国家科技部国际合作交流专项(2012DFB30070)
内蒙古自治区科技厅重大基础研究开放课题(20130902)
农业部公益性行业科研专项2010
2011年度项目(201003075)
关键词
蒙古马
肠道细菌
总DNA提取
方法比较
Mongolian horse
intestinal bacteria
total DNA extraction
comparison of methods