一种植物总RNA的快速提取方法

A Protocol for Rapid Isolation of Total RNA from Plant Tissues

为验证NaAc/无水乙醇沉淀法提取植物总RNA的质量,本研究对小麦不同组织及其他不同种属植物花的总RNA进行提取和检验,结果表明,该方法提取的植物总RNA完整性好、纯度高。以Tubulin为参照对其余4个持家基因GAPDH、Actin、Rubisco和Ubiquitin的表达情况进行分析。扩增结果表明,选定的各持家基因在叶锈菌侵染不同时间的小麦叶片中的表达量并不一致,且各基因在小麦叶片中的丰度也存在差异。通过对几种基因的扩增,进一步证实该方法能够满足一般分子生物学下游操作的要求,是一种理想的植物总RNA提取方法。

In order to check the quality of total RNA isolated by NaAc/ethanol precipitation,total RNA of radi- cle ,young leaves, young stems, immature seeds from wheat and flowers from different plant genus were extracted and checked. The results showed that total RNA obtained had good integrity and high purity. And the expression profiles of four frequently used house-keeping genes （ GAPDH,Actin, Rubisco, Ubiqutin） were analyzed by normalization with Tu- bulin gene. The results of the amplification showed that the transcripts of the selected house-keeping genes were differ- ent at different time points of wheat leaves inoculated by leaf rust pathogens, moreover, the copy numbers of each house-keeping gene were inconsistent in wheat leaves. PCR amplification shows that NaAc/ethanol precipitation can meet with demands of down stream experiment of common molecular biology and is an ideal method of plant total RNA extraction.