一种适用于RT-PCR的杉树类植物中总RNA提取的方法

An Improved Method Suited to Extract Total RNA Used to RT-PCR from Fir Plant

杉树类植物含大量的多糖、多酚类物质,这使得此植物的总RNA提取较困难。研究通过比较几种有代表性的提取植物总RNA的方法,建立了一种酸酚法和PEG8000沉淀法相结合的改良的CTAB法。此方法操作简单,使用的试剂均为常规试剂,成本低,能够有效去除植物组织中多糖和多酚类等次生物质,从而得到高质量的RNA。使用这种改良的方法制备的总RNA产物对P450基因成功地进行了反转录聚合酶链式反应(reversetrans-criptase-polymerasechainreaction,RT-PCR),证明此方法是一种适用于RT-PCR的杉树类植物组织中总RNA提取的方法。

Fir plants contain plenty of polysacchrides and polyphenols.Comparing several standard methods for extracting total RNA from fir plant, a modified method which can eliminate the interferences of polysacchrides and polyphenols in the plant tissues was constructed in the present study. The high-quality RNA sample obtained by the improved CTAB method used to RT-PCR (reverse transcriptase-polymerase chain reaction) of P450 gene study,and the result indicated that the method is easier for operation and is at lower cost.