一种从肠道微生物群中提取细菌DNA用于ERIC-PCR检测的简单快速方法。
A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection.
作者信息
Yang Jin-Long, Wang Ming-Shu, Cheng An-Chun, Pan Kang-Cheng, Li Chuan-Feng, Deng Shu-Xuan
机构信息
Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Yaan 625014, Sichuan Province, China.
出版信息
World J Gastroenterol. 2008 May 14;14(18):2872-6. doi: 10.3748/wjg.14.2872.
AIM
To develop a simple and convenient method for extracting genomic DNA from intestinal microflora for enterobacterial repetitive intergenic consensus (ERIC)-PCR detection.
METHODS
Five methods of extracting bacterial DNA, including Tris-EDTA buffer, chelex-100, ultrapure water, 2% sodium dodecyl sulfate and 10% Triton-100 with and without sonication, were compared with the commercial fecal DNA extraction kit method, which is considered as the gold standard for DNA extraction. The comparison was based on the yield and purity of DNA and the indexes of the structure and property of micro-organisms that were reflected by ERIC-PCR.
RESULTS
The yield and purity of DNA obtained by the chelex method was similar to that obtained with the fecal DNA kit. The ERIC-PCR results obtained for the DNA extracted by the chelex method and those obtained for DNA extracted with the fecal DNA kit were basically the same.
CONCLUSION
The chelex method is recommended for ERIC-PCR experiments in view of its simplicity and cost-effectiveness; and it is suitable for extracting total DNA from intestinal micro-organisms, particularly for handling a large number of samples.
目的
开发一种简单便捷的从肠道微生物群中提取基因组DNA的方法，用于肠杆菌重复基因间共识序列（ERIC）-PCR检测。
方法
将五种提取细菌DNA的方法，包括Tris-EDTA缓冲液法、螯合树脂100法、超纯水法、2%十二烷基硫酸钠法以及有超声处理和无超声处理的10% Triton-100法，与被视为DNA提取金标准的商业粪便DNA提取试剂盒法进行比较。比较基于DNA的产量和纯度以及ERIC-PCR所反映的微生物结构和特性指标。
结果
螯合树脂法获得的DNA产量和纯度与粪便DNA试剂盒法相似。螯合树脂法提取的DNA的ERIC-PCR结果与粪便DNA试剂盒法提取的DNA的ERIC-PCR结果基本相同。
结论
鉴于螯合树脂法简单且具有成本效益，推荐用于ERIC-PCR实验；它适用于从肠道微生物中提取总DNA，尤其适用于处理大量样本。
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