两种适用于葡萄不同组织RNA提取方法的比较

Comparison of Two Methods for RNA Extraction from Different Tissues of Grape(Vitis vinifera L.)

以不同葡萄组织为材料对目前常用的2种总RNA提取方法一改良SDS法和CTAB-LiCl法进行研究。2种RNA提取方法中均不使用酚。采用这两种方法从葡萄不同组织中均成功地提取到RNA，琼脂糖凝胶电泳结果显示28s和18SrRNA条带完整清晰。检测A260/A280 值分布在1．7-2．0之间，A260/A230值分布于1．9-2．3之间，说明RNA质量较高。管家基Actin和ACS5基因的检测表明2种方法所得RNA能够满足RT-PCR和基因克隆等研究需要。改良SDS法的RNA得率是CTAB-LiCl法的RNA得率的2～3倍，而CTAB-LiCl法获得的RNA纯度高。可以根据原料的数量和对RNA质量的要求来选取最佳提取方法。

Two total RNA extraction methods commonly used at present, modified SDS and CTAB-LiCl methods, were studied with different grape tissues. Phenol wasn＇t used in the two methods. Higher quality total RNAs from different grape tissues were obtained with both of two methods. Agrose gel electrophoresis showed that 28S and 18S bands were clear and bright. The ratio of A260/A280was from 1.7 to 2.0 while that of A260/A230 was from 1.9 to 2.3 which demonstrated the high quality of the isolated RNAs. The results of amplification of Actin, a housekeeping gene in plant, and ACS5 gene showed that the RNAs obtained could meet the requirements of RT- PCR and gene cloning. The yield of RNA obtained by modified SDS method was 2-3 times that by CTAB-LiC1 method. The total RNA isolated by the CTAB-LiCl method was highly integrate and pure. A preferable method could be selected according to the material quantity and the requirements of the RNA quality.