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Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components

Plant Molecular Biology Reporter volume 15, pages 8–15 (1997)Cite this article

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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Abbreviations

CTAB:

hexadecyltrimethylammonium bromide

EtOH:

ethanol

PVP:

polyvinylpyrrolidone

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Authors and Affiliations

  1. Agriculture Canada, Eastern Cereal and Oilseed Research Centre, K1A 0C6, Ottawa, Ontario, Canada

    Sue Porebski, L. Grant Bailey & Bernard R. Baum

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  1. Sue Porebski
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  2. L. Grant Bailey
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  3. Bernard R. Baum
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Porebski, S., Bailey, L.G. & Baum, B.R. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol Biol Rep 15, 8–15 (1997). https://doi.org/10.1007/BF02772108

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