甘薯块根的五种RNA提取方法的比较研究

Comparative Study on Five RNA Extraction Methods of Ipomoea batatas Root Tuber

为了确立甘薯块根总RNA最适提取方法,为富含色素、多糖、蛋白质等物质的试验材料的RNA提取提供参考。本研究选择紫薯1号、徐紫薯L7和浙薯1号的块根作为研究材料,采用Trizol法、RNA prep Pure Plant Kit法、RNA plant Plus法、研卉总RNA提取法和CTAB法共五种提取方法对甘薯块根总RNA进行提取,并考察各方法 RNA提取浓度和纯度。研究显示,Trizol法、RNA plant Plus法和RNA prep Pure Plant Kit法在甘薯块根总RNA提取中存在弊端,提取的RNA质量较差。CTAB法和研卉总RNA提取法提取的RNA质量较好,可有效去除各类杂质,且可用于反转录并扩增出IbTubA基因的特异条带,其中,CTAB法步骤比较繁琐但提取的RNA浓度高,而研卉总RNA提取法操作简便但提取的RNA浓度相对较低。

The aim of this study was to establish the optimal extraction method for total RNA from Ipomoea batatas （sweet potato） root tuber, so as to provide reference for RNA extraction of test materials that are rich in pigment, polysaccharide, protein and other substances. In this study, the root tuber of XZS 1, XZS L7 and ZS 1 cultivars were selected as the research materials, and the total RNA from the sweet potato root tubers were extracted by Trizol method, RNA prep pure Plant Kit method, RNA plant Plus method, Yanhui total RNA extraction method and CTAB method, and the concentration and purity of extracted RNA by each method were analyzed. The results showed that Trizol method, RNA plant Plus method and RNA prep Pure Plant Kit method had disadvantages in total RNA extraction of sweet potato root tuber, and the extracted RNA was of poor quality. The qualities of RNA that extracted by CTAB method and Yanhui total RNA extraction method were better than other methods. The two methods could effectively remove all kinds of impurities and could be used for the reverse transcription and the amplification of lbTubA gene specific bands. In addition, the steps of CTAB method were more complex but the extracted RNA was in high concentration, while the Yanhui total RNA extraction method was simple but the RNA concentration was relatively lower.