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.
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.
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2013年第3期439-442,共4页
Journal of South China Agricultural University
基金
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)
