Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components |
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Authors: | Sue Porebski L Grant Bailey Bernard R Baum |
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Institution: | (1) Agriculture Canada, Eastern Cereal and Oilseed Research Centre, K1A 0C6 Ottawa, Ontario, Canada |
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Abstract: | A relatively quick, inexpensive and consistent protocol for extraction of DNA from expanded leaf material containing large
quantities of polyphenols, tannins and polysaccharides is described. Mature strawberry leaves, which contain high levels of
these secondary components, were used as a study group. The method involves a modified CTAB extraction, employing high salt
concentrations to remove polysaccharides, the use of polyvinyl pyrrolidone (PVP) to remove polyphenols, an extended RNase
treatment and a phenol-chloroform extraction. Average yields range from 20 to 84 μg/g mature leaf tissue for both wild and
cultivated octoploid and diploidFragaria species. Results from 60 plants were examined, and were consistently amplifiable in the RAPD reaction with as little as 0.5
ng DNA per 25-μL reaction. Presently, this is the first procedure for the isolation of DNA from mature strawberry leaf tissue
that produces consistent results for a variety of different species, both octoploid and diploid, and is both stable and PCR
amplifiable before and after extended storage. This procedure may prove useful for other difficult species in the family Rosaceae. |
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Keywords: | Fragaria DNA extraction RAPD strawberry |
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