林蛙输卵管总RNA提取方法的优化及RT-PCR验证

Optimization of Total RNA Isolation fromRana chensinensis Oviduct and Validate the Method by RT-PCR

分别使用改良的十六烷基三甲基溴化铵(CTAB)法、TRIzol法和RNAiso Plus法对林蛙的输卵管组织进行总RNA提取,考察不同方法对总RNA提取量的影响.结果表明:利用改良的RNAiso Plus法并经进一步纯化,获得总RNA的A260/A280平均值为1.96,平均得率为56.1μg/g;通过反式多聚酶链式反应(RT-PCR)获得了林蛙GAPDH基因和EF-1-α基因的部分序列.

We compared the extraction effects of improved CTAB,TRIzol and RNAiso Plus,and investigated the extraction yield with different methods.It presents the advantages of high quality total RNA extracted by the optimized method,A260/A280ratio was 1.96and yield was more than 56.1μg/g.Through reverse transcription-polymerase chain reaction（RT-PCR）,we got GAPDHand EF-1-αsequences.