红麻叶片高质量RNA提取方法比较分析

Comparison and analysis of methods of extracting high-quality total RNA from kenaf leaves

红麻叶片中富含多糖、多酚和其他次生代谢物质,影响其RNA的产量和质量.本研究应用CTAB法、热硼酸法、SDS法、Trizol法和商品化试剂盒5种方法提取福红992幼嫩叶片总RNA,并对各方法提取效果进行比较.结果表明,热硼酸法效果最佳,其次是CTAB法和SDS法,而Trizol法、试剂盒提取效果不理想.采用热硼酸法提取的红麻叶片总RNA能够成功进行反转录和制备cDNA.

RNA isolation from kenaf leaves is difficult due to containing large amounts of polysaccharides,polyphenol and other secondary metabolites,affecting both the quality and quantity of RNA.In this study,total RNA from the leaves of Fuhong 992 was extracted by the five methods of CTAB,hot-borate,SDS,Trizol and Biotake corporation kit,carried on the extraction effect comparative analysis.The results showed that the method of hot-borate was more suitable to isolate total RNA from kenaf leaves,next were CTAB and SDS these two methods,Trizol and Biotake corporation kit did not obtain the ideal result.It is capable of obtaining high quality and high integrity total RNA from kenaf leaves using hot-borate method,and conducting its reverse transcription to prepare cDNA.It demonstrated that the total RNA isolated by this method was of sufficient quality for subsequent molecular applications.