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A RAPID DNA EXTRACTION METHOD IS SUCCESSFULLY APPLIED TO ITS-RFLP ANALYSIS OF MYCORRHIZAL ROOT TIPS
Citation:
Sahalie, J., K. J. Martin, AND P T. Rygiewicz. A RAPID DNA EXTRACTION METHOD IS SUCCESSFULLY APPLIED TO ITS-RFLP ANALYSIS OF MYCORRHIZAL ROOT TIPS. Presented at Oregon State University Center for Gene Research and Biotechnology Fall Retreat, Newport, OR, September 29, 2000.
Description:
A rapid method for extracting DNA from intact, single root tips using a Xanthine solution was developed to handle very large numbers of analyses of ectomycorrhizas. By using an extraction without grinding we have attempted to bias the extraction towards the fungal DNA in the mantle over plant DNA internal to the tip. Xanthogenic acid was used to dissolve the polysaccharides in the cell walls and a mild detergent weakened the lipid membranes. Single, lyophilized root tips were briefly sonicated in Xanthine solution and incubated on an orbital shaker (60 C). The solution was then transferred to a new tube for PEG precipitation. After two ethanol rinses the DNA was dried and redissolved for use in PCR. Large numbers of tips can be processed in this way to determine PCR-RFLP patterns for ectomycorrhizas at a scale advantageous for ecological studies.