Genomic DNA isolation from Hippophaë rhamnoides subsp. carpatica Rousi for RAPD fingerprinting

The assessment of the genetic diversity of Hippophae rhamnoides L. species became an important prerequisite for its conservation and marker assisted breeding. The first step in this type of studies is represented by the optimization of genomic DNA extraction in order to get high quality DNA, suitable for generation of molecular markers, such as RAPD. Thus, a variety of extraction methods have been evaluated for their efficiency and it was noticed that the quantity and quality of isolated DNA varied considerably among the various extraction protocols. SDS protocol and the commercial kit Wizard Genomic DNA purification represented the best DNA extraction methods for sea buckthorn leaf tissue, when an enzymatic digestion prior to isolation using CelluACE TM XG System kit was performed. For further RAPD and other PCR-based methods the most suitable DNA extraction is SDS + CelluACE TM XG System, due to the quality of obtained band pattern.