羊蹄甲果荚中总RNA提取的新方法

New Method for Total RNA Extraction in Legume of Bauhinia Variegata

高质量RNA的获得是开展羊蹄甲分子生物学研究的基础,但要获得高质量的植物总RNA比较困难,RNA的提取方法会因植物材料的不同而有较大差异。目前已有多种从植物组织中提取RNA的方法。但是由于羊蹄甲中的次生物质含量较高,这些常用方法并不适用于羊蹄甲RNA的提取。本文介绍一种羊蹄甲果荚中总RNA的提取方法,它可以有效的排除羊蹄甲组织中大量存在的多酚类和多糖类物质的干扰。所得RNA样品经紫外分光光度计检测和1%琼脂糖凝胶电泳分析证明无明显降解,且具有较高的纯度。获得的RNA样品在纯度和浓度上都可以满足RT-PCR,Northernblot和cDNA文库构建等分子生物学实验的要求。本方法尤其适用于多酚和多糖含量较高的植物组织的RNA提取,且试剂简单经济,试验结果稳定,重现性强。

The acquisition of high quality RNA is foundation of launching molecule biological study of Bauhinia variegata. But it is relatively difficult to extract quality total RNA those extracting methods very depending on different plant materials. Many methods for the extraction of RNA from plant tissues have been developed at present. Because the higher levels of the secondary compounds in Bauhinia variegata, some methods that usually used are not appropriate for the extraction of RNA from Bauhinia variegata tissues. A simple extraction method for the total RNA ofBauhinia variegata legume is introduced here. It can efficiently eliminate the interferent of polyphenolies and polysaeeharides which are very rich in Bauhinia variegata. The results of 1% agarose gel eleetrophoresis and ultraviolet speetrophotometer analysis show that the total RNA of Bauhinia variegata extracted through this method has no obvious degradation and has a good purity. The purity and concentration of extracted total RNA con meet the demand of molecular biology experiment such as RT-PCR, Northern blot experiment and eDNA library construction. It is especially useful for the RNA extraction of plant tissues which usually has plenty of polyphenolies and polysaeeharides. It is simple, inexpensive, stable and easily repeated.