一种快速、高效的橡胶树胶乳总RNA 提取方法

胶乳是橡胶树(Hevea brasiliensis)乳管中特殊的细胞质, 主要由橡胶粒子、黄色体、F-W粒子和普通细胞质成分构成, 其中橡胶粒子占20%-40%, 蛋白含量高达1%-2%。由于高比例橡胶粒子和蛋白质的干扰, 目前使用的胶乳RNA提取方 法都具有步骤繁琐、胶乳需求量大、操作技巧性强不易掌握等缺点。为快速、高效地获取高质量的胶乳RNA, 我们在现有方法的基础上摸索出一套步骤简单、容易操作、快速、高效提取橡胶树胶乳总RNA的简易方法, 获得了较好的实验效果。紫外分光光度计、RT-PCR和RACE分析结果表明, 使用该方法提取的胶乳RNA质量完全能够满足相应的分子操作, 但所需时间仅为目前常用方法的50%, RNA获得率提高了2-3倍, 操作难度大大降低。

A Rapid and Efficient Protocol for Total RNA Isolation from Latex of Hevea brasiliensis

Latex is a special cytoplasm of laticifer in Hevea brasiliensis and is mainly composed of rubber particles, lutoids, FW particles and all components of normal cytoplasm; rubber particles take up 20%-40% and total protein 1%-2%. Because of the high protein and rubber-particle content of latex, methods for total RNA isolation of latex, modified from Kush‘s protocol, contain problems such as complicated processes, difficult operation and large demand for latex. Here, we describe a rapid and efficient protocol for total RNA isolation from latex, which cut the process time by 50% and increased the harvest of total RNA by 2-3 times. Analysis by ultraviolet spectrophotometry and RT-PCR showed the isolated total RNA with good quality for further molecular experiments.