海岸植物许树和单叶蔓荆提取total RNA的简便方法

目的]介绍海岸植物叶片组织total RNA提取的一种简单、便捷方法。方法]取800~1 000 mg许树和单叶蔓荆的鲜叶,加液氮研磨后制成提取液,提取液加试剂4℃下离心,冰浴后室温下离心,依次添加试剂后室温下离心,即得total RNA样品。结果]分光光度计测试提取的total RNA的A260/A230大于2.10,A260/A280在1.93左右;RNA溶液中RNA浓度约为4.0μg/μl。该方法用了较少试剂种类;每根柱只需洗4次便可获得较好质量的total RNA;一次研磨800~1000 mg材料可提取约700μg total RNA;电泳检查表明提取的totalRNA符合分子生物学研究要求,可直接用于各种分子生物学试验。结论]该方法是一种以较少试剂、较小费用、短时间内提取较高质量与产量的海岸植物叶片组织total RNA的简单、便捷方法。

A Simple and Convenient Method of Extracting Total RNA from Coastal Plants Clerodendron inerme and Vitex rotundifolia

Objective] The purpose was to introduce a simple and convenient method of extracting total RNA from leaf tissues of coastal plants.Method] The extracts was prepared by grinding 800-1 000 mg fresh leaves of Clerodendron inerme and Vitex rotundifolia added with liquid nitrogen.The sample of total RNA was prepared by centrifugating the extracts added with reagent at 4 ℃,centrifugating at room temperature after ice bath and centrifugating at room temperature after adding reagent in order.Result] A260/A230 of the extracted total RNA determined by spectrophotometer was bigger than 2.10,with A260/A280 of about 1.93.The RNA content in the solution was about 4.0 μg/μl.Fewer kinds of reagents were used by this method,the total RNA with better quality could be obtained by washing each column for only 4 times,and about 700 μg total RNA could be extracted by grinding 800-1 000 mg materials for once.The electrophoretic check showed that the extracted total RNA accorded the requirements of molecular biological research and could be directly used in various molecular biological experiments.Conclusion] This method was a simple and convenient method of extracting total RNA with higher quality and output from leaf tissues of coastal plants in short time,with less agent and cost.