Fang G, Hammar S, Grumet R
Department of Horticulture, Michigan State University, East Lansing 48824.
Biotechniques. 1992 Jul;13(1):52-4, 56.
A quick and inexpensive method has been demonstrated to remove polysaccharide contamination from plant DNA. Isolated plant genomic DNA with polysaccharide contaminants was dissolved in TE (10 mM Tris-HCl, pH 7.4, 1 mM EDTA) with NaCl ranging from 0.5-3.0 M, then precipitated with two volumes of ethanol. Most of the polysaccharides were removed effectively in a single high-salt precipitation at 1.0-2.5 M NaCl. At 3.0 M NaCl, the salt precipitated out of solution. Purified DNA was easily digested by either HindIII or EcoRI and was satisfactory as a template for PCR. The results show that high-salt precipitation effectively removed polysaccharides and their inhibitory effects on restriction enzyme and Taq polymerase activity.
已证明一种快速且廉价的方法可去除植物DNA中的多糖污染。将含有多糖污染物的分离植物基因组DNA溶解于含0.5 - 3.0 M NaCl的TE（10 mM Tris - HCl，pH 7.4，1 mM EDTA）中，然后用两倍体积的乙醇沉淀。在1.0 - 2.5 M NaCl的单次高盐沉淀中，大部分多糖被有效去除。在3.0 M NaCl时，盐从溶液中析出。纯化后的DNA很容易被HindIII或EcoRI消化，并且作为PCR模板令人满意。结果表明，高盐沉淀有效地去除了多糖及其对限制酶和Taq聚合酶活性的抑制作用。
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