Genomic DNA Extraction for Authentication of Clematis florida Thunb. var. plena D. Don

  Objective   A rapid and economical method to extract the genomic DNA from Clematis florida Thunb.var. plena D. Don for commercial authentication of the valuable Chinese herbal medicine was established.

  Method  CTAB, Testing Kit, SDS, and the high salt/low pH methods, along with a modified CTAB method, were applied to extract genomic DNA from leaves of the herbal plant. The quality and integrity of the extracted DNA samples were compared using agarose gel electrophoresis. Then, by bidirectional sequencing, the selected molecular pharmacognosy method was used to differentiate between the authentic material and a counterfeit for methodology verification.

  Result  The modified CTAB method yielded high purity DNA with clear and bright bands on SSR primer and ITS2 sequence amplifications. The PCR amplified products of the extracted DNA were used in the bidirectional sequencing to correctly identify and accurately differentiate the true and faux products.

  Conclusion  The improved CTAB method established in this study could effectively extract the genomic DNA of the target plant material. It was simple, rapid and reliable in extracting high quality DNA for molecular pharmacognosy identification on Clematis florida Thunb. var. plena D. Don.