A quick and inexpensive method for removing polysaccharides from plant genomic DNA.
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.
Fang
G
G
Department of Horticulture, Michigan State University, East Lansing 48824.
Hammar
S
S
Grumet
R
R
D004247:DNA / Q000302:isolation & purification*; D000431:Ethanol; D005821:Genetic Techniques / Q000191:economics; D010944:Plants / Q000235:genetics*; D011134:Polysaccharides / Q000302:isolation & purification*; D013997:Time Factors
D011134:Polysaccharides; D000431:Ethanol; D004247:DNA