A simple technique for removing plant polysaccharide contaminants from DNA.
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.
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Plant Biotechnology Center, Baylor University, Waco, TX 76798.
Adams
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D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D005069:Evaluation Studies as Topic; D005821:Genetic Techniques; D010944:Plants / Q000032:analysis*; D011134:Polysaccharides / Q000302:isolation & purification* / Q000378:metabolism
D011134:Polysaccharides; D004247:DNA; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII