岩白菜幼叶总RNA的提取方法研究

Study on the Total RNA Extraction from Young Leaves of Bergenia purpurascens

为了筛选出提取岩白菜幼叶总RNA的最佳方法,为岩白菜的转录组测序提供技术支撑,本文采用6种试剂盒法提取岩白菜幼叶的总RNA,通过凝胶电泳和BIO-RAD核酸蛋白检测仪检测提取的RNA样品的质量和纯度,并对提取效果进行了分析。结果表明:用Trizol Kit、Trizol A^+Kit、Transzol Up Kit提取的幼叶总RNA没有28S rRNA和18S rRNA条带;用inun PREP Plant RNA Kit提取的RNA具有28S rRNA和18S rRNA条带,但条带模糊、含量低;用Transzol Plant Kit和Easy Pure Plant RNA Kit提取的RNA具有28S rRNA和18S rRNA这2条清晰的条带,但后者提取的总RNA的得率和纯度更高。综合考虑后认为Easy Pure Plant RNA Kit法是快速提取岩白菜幼叶总RNA的最佳方法。

In order to screen out the best method of extracting total RNA from young leaves of Bergenia purpurascens and provide technique support for RNA-sequencing,six reagent kits such as Trizol kit,Trizol A~＋Kit,Transzol Plant Kit,Transzol Up Kit,Easy Pure Plant RNA Kit and inun PREP Plant RNA Kit were used and the quality and purity of the total RNAs extracted with them were tested by gel electrophoresis and BIO-RAD Smart Spec Plus. The results showed that RNAs extracted with Trizol Kit、Trizol A~＋Kit and Transzol Up Kit lacked the bands of 28 S rRNA and 18 S rRNA. The RNA extracted with inun PREP Plant RNA Kit showed two weak bands of 28 S rRNA and 18 S rRNA. The RNAs extracted by Transzol Plant and Easy Pure Plant RNA Kit exhibited two bright bands of 28 S rRNA and 18 S rRNA,but the purity and yield rate of the latter were better. Extraction with Easy Pure Plant RNA Kit was the best method to extract the total RNA from young leaves of B. purpurascens.