Manning K
Department of Plant Physiology, Institute of Horticultural Research, West Sussex, England.
Anal Biochem. 1991 May 15;195(1):45-50. doi: 10.1016/0003-2697(91)90292-2.
The purification of nucleic acids from plant tissue is often made difficult by the presence of contaminating carbohydrate polymers and polyphenols. A procedure for the simultaneous isolation of DNA and translatable RNA from plants is described. The method removes most of the polysaccharides and polyphenols extracted with nucleic acids in a single step by taking advantage of differences in solubility of these compounds in the solvent 2-butoxyethanol. Stepwise addition of 2-butoxyethanol to phenol extracts of specific ionic strength precipitates nucleic acids largely free of contaminants. Subsequent separation of RNA from DNA by precipitation with LiCl was optimised to give a high recovery of translationally active RNA. Successful isolation of nucleic acids from strawberry (Fragaria X ananassa) receptacle, a particularly recalcitrant tissue, and from a range of tissues of other plant species demonstrates the general applicability of the method.
植物组织中核酸的纯化常常因存在污染性的碳水化合物聚合物和多酚而变得困难。本文描述了一种从植物中同时分离DNA和可翻译RNA的方法。该方法利用这些化合物在溶剂2-丁氧基乙醇中的溶解度差异,在一步操作中去除了大部分与核酸一起提取的多糖和多酚。向特定离子强度的酚提取物中逐步添加2-丁氧基乙醇,可沉淀出基本不含污染物的核酸。随后通过用LiCl沉淀从DNA中分离RNA的步骤得到了优化,以实现翻译活性RNA的高回收率。从草莓(Fragaria X ananassa)花托(一种特别难处理的组织)以及其他植物物种的一系列组织中成功分离出核酸,证明了该方法的普遍适用性。