Comparison of Protocols for DNA Extraction from Cannabis sativa seeds

Salvador SOLER, Alicia SIFRES, Ernesto R LLOSA, Matías LLAMAS, Santiago VILANOVA, Jaime PROHENS

Abstract


Cannabis sativa is an important crop cultivated both for industrial (var. sativa) and medicinal and recreational (var. indica) purposes. Cultivation of var. indica plants is generally forbidden and this difficults genetic studies. An alternative is the extraction of DNA from seed embryos. In order to develop an efficient protocol for DNA extraction of C. sativa seeds we have tested six DNA extraction methods in seeds and leaves of a C. sativa var. sativa accession. We found that the best protocol is the CTAB-modified with phenol:chlorophorm:isoamyl washing, which allowed a large quantity of high quality DNA per seed. This method was tested in seeds of three C. sativa var. indica accessions and seven SSR markers tested could be amplified successfully. In summary, we have developed a highly efficient method for DNA extraction of individual seeds of C. sativa. This method will be useful for genetic studies in this species.

Keywords


CTAB; embryos; extraction kits; molecular markers; SSRs

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DOI: http://dx.doi.org/10.15835/buasvmcn-hort:9255




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