大豆子叶RNA和DNA的同步抽提及质量分析

Simultaneous Isolation of RNA and DNA from Soybean Cotyledon and Analysis of Their Quality

本文旨在建立一种适用于大豆子叶的RNA和DNA同步抽提方法,得到高质量的核酸以满足后续实验要求。该方法先用异硫氰酸胍裂解大豆子叶,再用常规苯酚/氯仿抽提去除杂质,最后用LiCl选择性沉淀核酸。核酸的完整性和纯度检测表明抽提的总RNA和DNA质量良好,进一步的RT-PCR和PCR检测显示抽提的总RNA和DNA质量适合后续的实验要求。

A method for simultaneous isolation of RNA and DNA is desirable when a limited amount of sample is available. The purpose of this paper was to present a new method to isolate DNA and RNA simultaneously from soybean cotyledon （Glycine max）. Sample was first lysed and homogenized in liquid nitrogen and the presence of a guanidine isothiocyanate-containing buffer. The lysed mixture was then extracted by a phenol/chloroform mixture e- quilibrated with Tfis-HC1 buffer. The nucleic acids （DNA and RNA） were differentially precipitated by the addition of LiC1 and ethanol. The isolated nucleic acids had a good integrity and purity and were suited for the further experiments.