不同肠球菌基因组DNA提取方法对PCR鉴定的影响

为研究肠球菌基因组DNA的不同提取方法对16SrRNA基因序列PCR、种属多重PCR、RAPDPCR等肠球菌鉴定结果的影响,分别采用6种基因组DNA提取方法(水煮法1、水煮法2、常规酚/氯仿抽提法、chelex-100法1、chelex-100法2和试剂盒法),对1株肠球菌标准菌株ATCC29212基因组DNA进行提取,以这些分别提取的DNA为模板在相同的扩增条件下进行PCR检测.结果表明,在16SrRNA基因序列PCR和种属多重PCR检测中以水煮法1进行肠球菌DNA的提取最适宜,而试剂盒法在RAPDPCR检测中效果最佳.

Influences of different methods of extracting enterococccal genome DNA on PCR identification

In order to study the effect of different methods of extracting DNA from Enterococcus on the identifying result through 16S rRNA gene sequence PCR,genus multiplex PCR and RAPD PCR,this research used six methods of extracting genome DNA i.e.the water boiling 1 method,the water boiling 2 method,the conventional phenol-chloroform method,the chelex-100 1 method,the chelex-100 2 method and the genomic DNA kit method to extract the genome DNA of a standard Enterococcus termed ATCC29212,then the DNA samples undre amplified with the templates under the same condition.The results showed that the water boiling 1 method was the most suitable method to extract the DNA in the 16S rRNA gene sequence PCR and genus multiplex PCR,and the genomic DNA kit was the best one to extract the DNA in the RAPD PCR.