Abstract:

To investigate the genetic diversity and identification of germplasm resources, the genomic DNA was extracted from the leaves of Tussilago farfara L. by using the SDS, CTAB, modified SDS and modified CTAB methods. The extraction effects of different methods were compared. The subsequent ISSR-PCR condition was explored, using the DNA harvested from modified CTAB methods. The results showed that the modified SDS and CTAB methods could effectively obtain high quality DNA, and the most suitable mixture was as follows: total 20μl reaction volume containing template DNA 20ng, 1.0 UTaq DNA polymerase, 2.00 mmol/L Mg2+, 0.20 mmol/L dNTPs, 0.6 μmol/L pimer, 1×PCR buffer.