用于PCR扩增的细菌DNA提取方法比较
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国家自然科学基金(31070103,31200006) ;广东省自然科学基金重点项目( 10251007002000001);广东省科技计划项目(2009B030802013)


A Comparative Study on Bacteria DNA Extraction Methods Used for PCR Amplification
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    摘要:

    以14个属的革兰阴性菌和阳性菌为研究对象,采用16S rRNA基因PCR扩增对4种细菌DNA提取法进行了比较.结果表明,对细菌DNA的提取效果排序,依次为冻融法、煮沸法、碱解法、ROSE法.冻融法效果最好,具有成本低、省时省力、无污染等优点,但仍存在少数革兰阳性菌DNA提取失败的现象.因此,在菌株较多的情况下可先采用冻融法提取细菌DNA,对少数提取失败的菌株,弃冻融法得到的上清液,再以SDS法、CTAB法或DNA试剂盒的提取进行补充.采用该操作流程既能节省成本和时间,又减少了提取过程中有机废弃物的产生.

    Abstract:

    The 16S rRNA genes amplification effects of the four simple total DNA extraction methods (freeze-thaw, boiling, alkaline lysis, and ROSE) were compared by extracting DNA from the Gram-negative and gram-positive bacterial strains of 14 genera. The results showed that the extraction effects of the methods from superior to inferior were in the following order: freeze-thaw, boiling, alkaline lysis and ROSE. The freeze-thaw method had advantages of cost-effective, time-and labor-saving and non-pollution. However, it was not always effective for minority gram-positive bacteria. When the DNA extraction of a large number of bacterial strains was required, the freeze-thaw method to extract total DNA of most bacteria may be first used. For the minority of DNA extraction samples which failed, it was advisable to discard the suspension from the freeze-thaw method, then use the chemical methods such as CTAB, SDS or DNA extraction kit for supplement. The freeze-thaw method to extract total DNA could not only save time and reduce cost, but also decrease the production of organic waste during DNA extrction.

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冯广达,陈美标,羊宋贞,朱红惠.用于PCR扩增的细菌DNA提取方法比较[J].华南农业大学学报,2013,34(3):439-442

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  • 收稿日期:2012-09-03
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  • 在线发布日期: 2013-06-27