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为了筛选三峡库区草坪草狗牙根(Cynodon dactylon)叶片RNA的提取方法,试验采用RNAprep pure植物总RNA提取试剂盒进行狗牙根的RNA提取。结果表明:在RNA提取试剂盒裂解液中加入1%β-巯基乙醇,提取的RNA完整性和纯度都较好,可以清晰辨别出28S和18S条带,5S条带较淡;生物分光光度计测得该方法提取的RNA浓度为6.85μg/mL,其OD260/OD280和OD260/OD230值分别为1.89和2.18,没有DNA和蛋白质污染,达到了高质量RNA样品的标准;RT-PCR提取的RNA反转录后可以满足后续分子生物学试验的要求。说明在RNAprep pure植物总RNA提取试剂盒的裂解液中加入1%β-巯基乙醇可以提取到高质量的狗牙根叶片RNA。
In order to screen the RNA extracted from leaves of lawn grass Cynodon dactylon in the Three Gorges Reservoir Area, RNA extraction was performed using RNAprep pure total RNA extraction kit. The results showed that 1% β-mercaptoethanol was added into the lysis buffer of RNA extraction kit, the integrity and purity of the extracted RNA were good, the 28S and 18S bands could be distinguished clearly, and the 5S bands were lighter. The biological spectrophotometer The concentration of RNA extracted by this method was 6.85μg / mL and the OD260 / OD280 and OD260 / OD230 values were 1.89 and 2.18, respectively. There was no DNA and protein contamination and the standard of high quality RNA samples was reached. The RNA extracted by RT-PCR After reverse transcription to meet the requirements of subsequent molecular biology experiments. This indicated that 1% β-mercaptoethanol was added to the RNAprep pure plant RNA extraction kit to extract high quality bermudagrass leaf RNA.