Comparison of Protocols for DNA Extraction from Cannabis sativa seeds

  • Salvador SOLER Universitat Politècnica de València
  • Alicia SIFRES Universitat Politècnica de València
  • Ernesto R LLOSA Hemp Trading SLU
  • Matías LLAMAS Maris Seed Spanish
  • Santiago VILANOVA Universitat Politècnica de València
  • Jaime PROHENS Universitat Politècnica de València
Keywords: CTAB, embryos, extraction kits, molecular markers, SSRs


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.