适宜西瓜mRNA差异显示分析的几种总RNA提取方法的比较

以西瓜花蕾为材料,初步建立了mRNA差异显示技术体系。利用改进的SDS/酚法、改进的异硫氰酸胍法和BIOZOL试剂提取法提取西瓜花蕾总RNA,比较RNA产率、纯度和电泳图谱等分析来确立适于西瓜花蕾RNA分离的方法。研究结果表明,改进的异硫氰酸胍法提取的RNA呈现28S rRNA,18S rRNA和5S rRNA 3条较清晰的条带,其A260/A280值可达1.944,具有很高的纯度。用异硫氰酸胍法提取的总RNA进行差异显示分析表明,可得到理想的扩增片段。

Comparison of the Methods of RNA Isolation for mRNA Differential Display in Watermelon

The optimal mRNA Differential Display system in watermelon was established by using flower buds of watermelon as experimental materials.The authors isolated total RNA from buds of watermelon by the methods of SDS/phenol,Guanidine Thiocyanate and BIOZOL regent and established the method of RNA isolation suitable for buds of watermelon through the rate of yield,purity and electrophoresis of RNA.The results showed that there were three clear bands-28S rRNA,18S rRNA and 5S rRNA respectively-and little RNA was degraded by the method of Guanidine Thiocyanate,its ratio of A_(260)/ A_(280) was 1.944.Data of mRNA differential display showed that there were ideal amplified bands using the RNA isolated by the method of Guanidine Thiocyanate.