Kanai Masatake, Mano Shoji, Nishimura Mikio
Laboratory of Biological Diversity, Department of Evolutionary Biology and Biodiversity, National Institute for Basic Biology.
Laboratory of Biological Diversity, Department of Evolutionary Biology and Biodiversity, National Institute for Basic Biology; Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies);
J Vis Exp. 2017 Jan 11(119):55008. doi: 10.3791/55008.
Plant seeds accumulate large amounts of storage reserves comprising biodegradable organic matter. Humans rely on seed storage reserves for food and as industrial materials. Gene expression profiles are powerful tools for investigating metabolic regulation in plant cells. Therefore, detailed, accurate gene expression profiles during seed development are required for crop breeding. Acquiring highly purified RNA is essential for producing these profiles. Efficient methods are needed to isolate highly purified RNA from seeds. Here, we describe a method for isolating RNA from seeds containing large amounts of oils, proteins, and polyphenols, which have inhibitory effects on high-purity RNA isolation. Our method enables highly purified RNA to be obtained from seeds without the use of phenol, chloroform, or additional processes for RNA purification. This method is applicable to Arabidopsis, rapeseed, and soybean seeds. Our method will be useful for monitoring the expression patterns of low level transcripts in developing and mature seeds.
植物种子积累了大量由可生物降解有机物组成的储存储备。人类依靠种子储存储备作为食物和工业原料。基因表达谱是研究植物细胞代谢调控的有力工具。因此，作物育种需要种子发育过程中详细、准确的基因表达谱。获得高度纯化的RNA对于生成这些谱至关重要。需要有效的方法从种子中分离出高度纯化的RNA。在这里，我们描述了一种从含有大量油脂、蛋白质和多酚的种子中分离RNA的方法，这些物质对高纯度RNA的分离具有抑制作用。我们的方法能够在不使用苯酚、氯仿或额外RNA纯化过程的情况下从种子中获得高度纯化的RNA。该方法适用于拟南芥、油菜籽和大豆种子。我们的方法将有助于监测发育中和成熟种子中低水平转录本的表达模式。
