摘要
探索食品检测中快速高效提取革兰氏阳性菌基因组DNA的新方法。采用改良交替冻融法、CTAB法、苯酚氯仿法及试剂盒法等4种不同的方法提取金黄色葡萄球菌(Staphylococcus aureus)、单核细胞增生李斯特氏菌(Listeria monocytogenes)以及芽孢杆菌(Bacillus sp)的基因组DNA,并进行琼脂糖凝胶电泳检测及16S rDNA扩增检测。结果显示,以改良交替冻融法提取的基因组DNA为模板,可灵敏有效的扩增16S rDNA基因。该方法用时短,操作简单,成本低,能在短时间内完成大量样品的处理,满足普通的PCR扩增反应。
The aim was to research a new method for preparing genomic DNA from Gram-positive bacteria quickly and efficiently in food inspection. Four different methods including improved alternating freeze-thaw method, CTAB method, phenol-chloroform method and kit methodology were used for the DNA extraction of Staphylococcus aureus, Listeria monocytogenes and Bacillus sp. The agarose gel electrophoresis and 16S rDNA amplification were carried out for the DNA detection. The result indicated that the DNA prepared by the improved method was sufficiently pure for PCR. This method saves time and cost, practices easily as well, and valuable for rapid isolation of DNA from generous samples to meet in an ordinary PCR reaction.
出处
《食品工业》
北大核心
2011年第12期104-107,共4页
The Food Industry
基金
山东省科技攻关计划项目(2010GGC10522)
山东主要海产品微生物污染及其关键控制技术研究
关键词
食品检测
革兰氏阳性菌
DNA提取
PCR
food inspection
Gram-positive bacteria
DNA extraction
polymerase chain reaction
