一种从DNA中去除植物多糖污染物的简单技术。

A simple technique for removing plant polysaccharide contaminants from DNA.

作者信息

Do N, Adams R P

机构信息

Plant Biotechnology Center, Baylor University, Waco, TX 76798.

出版信息

Biotechniques. 1991 Feb;10(2):162, 164, 166.

PMID:2059438

Abstract

A survey of the inhibitory effects of various plant polysaccharides on DNA restrictions (HindIII and EcoRI) revealed that neutral polysaccharides (arabino-galactan, dextran, gum guar, gum locust bean, beta-glucan, inulin, laminaran, mannan and starch) were not very inhibitory. In contrast, acidic polysaccharides (carrageenan, dextran sulfate, gum ghatti, gum karaya, pectin and xylan) were very inhibitory, even at low concentrations. The Elutip-d (RPC-5 type resin) was evaluated for removal of the inhibitory polysaccharides. Used alone or in combination with a phenol/chloroform wash, it proved effective in removing the polysaccharide so that HindIII digestion was possible, except in the cases of carrageenan and dextran sulfate. In addition, the genomic DNA extracts from live oak (Quercus virginiana) and magnolia (Magnolia grandiflora) were sufficiently purified so that the DNAs could be restricted with both EcoRI and HindIII.

摘要

一项关于各种植物多糖对DNA限制性内切酶（HindIII和EcoRI）抑制作用的调查显示，中性多糖（阿拉伯半乳聚糖、葡聚糖、瓜尔豆胶、刺槐豆胶、β-葡聚糖、菊粉、海带多糖、甘露聚糖和淀粉）的抑制作用不太明显。相比之下，酸性多糖（角叉菜胶、硫酸葡聚糖、吉蒂胶、刺梧桐树胶、果胶和木聚糖）即使在低浓度下也具有很强的抑制作用。对Elutip-d（RPC-5型树脂）去除抑制性多糖的效果进行了评估。单独使用或与酚/氯仿洗涤联合使用时，它被证明能有效去除多糖，从而使HindIII消化成为可能，但角叉菜胶和硫酸葡聚糖除外。此外，从活橡树（弗吉尼亚栎）和木兰（大花木兰）中提取的基因组DNA得到了充分纯化，使得这些DNA能够被EcoRI和HindIII限制性内切酶切割。