摘要
采用TRIZOL试剂盒法、RNAsio试剂法和改良CTAB法提取香蕉叶片总RNA,并通过凝胶电泳、紫外分光光度法检测提取的RNA样品的质量。研究结果表明:改良CTAB法提取的RNA具有28SrRNA和18SrRNA两条清晰的条带,OD260/OD280在1.80～1.95之间,OD260/OD230在1.75～2.10之间,具有较高的纯度;其他两种方法获得的RNA纯度不够,降解严重。将提取的RNA分别逆转录成cDNA,经RT-PCR扩增,只有改良CTAB法出现清晰的条带,进一步证明改良CTAB法提取的RNA具有很高的纯度,可以满足进一步分子生物学研究的要求。
Total RNA was isolated from wild banana leaves by using the methods of TRIZOL Regent kit, RNAsio Regent and improved CTAB. The quality of total RNA was analyzed through gel electrophoresis and UV spectrometer. The results indicated that the RNA isolated by the method of improved CTAB showed clear bands of 28SrRNA and 18SrRNA, and the value of OD260/OD280 was 1.80 to 1.95, the value of OD260/OD230 was 1.75 to 2.10. RNA isolated by the other two methods degraded seriously. RNA could be reverse to cDNA. The cDNA was used for RT-PCR amplifying, but the electrophoresis figure by RT-PCR showed that there was one distinct band using the RNA temples isolated by the method of CTAB. These results demonstrated that the quality and purity of the RNA obtained by the method of improved CTAB could meet the demands of molecular biology experiment.
出处
《中国农学通报》
CSCD
北大核心
2009年第14期51-54,共4页
Chinese Agricultural Science Bulletin
基金
国家科技支撑计划"闽台特色果树的离体试管保存
香蕉抗冻蛋白基因克隆及转基因"(2007BAD07B01)
福建省亚热带果树及特种经济作物种质资源共享平台资助项目部分内容(2008N2001)
