新型试剂-D.S.G在烟草RNA提取与纯化中的应用

Application of New Reagent-D.S.G to Tobacco RNA Extraction and Purification

核酸提取是分子生物学实验的基础,提取高质量的核糖核酸(RNA)是构建cDNA文库和进行基因表达研究工作的前提。笔者比较了TRIREAGEN法、D.S.G法、异硫氰酸胍氯化铯密度梯度分离法、D.S.G和TRIREAGENT结合法、TRIREAGENT和D.S.G结合法等5种烟草RNA提取方法,结果表明D.S.G和TRIREAGENT两种试剂的配合使用均可有效地去除烟草植物细胞壁中的多聚糖及其它成份,获取较高质量的RNA。这说明新型试剂-D.S.G与TRIREAGENT的配合使用有利于获得高质量的RNA,但D.S.G单独使用效果并不佳。

The extraction of nucleic acid is the foundation in molecular biological experiments, and RNA with high quality is the premise for cDNA library construction and genes expression. Five methods of RNA extraction, TRIREAGENT, D.S.G, guanidinium isothiocyanate and cesium chloride density gradient centrifugation, D.S.G and TRIREAGENT, were compared. The method of TRIREAGENT and D.S.G showed that applying the later two methods to extract tobacco RNA could wipe off the polysaccharide and other components of the tobacco cell wall. The result indicated that combination of D.S.G and TRIREAGENT was propitious extraction method of tobacco RNA with high quality. However, RNA quality wasn＇t improved by D.S.G only.