Abstract:

The purpose of this study was to obtain the best method for extracting genomic DNA from Taraxacum kok-saghyz Rodin and to establish the best reaction condition of the RAPD amplification system. The high quality genomic DNA of Taraxacum kok-saghyz Rodin could be obtained from young leaves by the improved CTAB method. Some reaction conditions of PCR for RAPD were optimized by setting the gradient with template DNA concentration, primer concentration, dNTPs concentration, Taq DNA Polymerase concentration and Mg2+ concentration through a single factor experiment. And the optimized reaction system and program of RAPD were as follows. The reactions were performed in a volume of 20 μL, which containing 30 ng DNA template、1.75U Taq DNA polymerase、0.25 mmol/ L dNTPs、2.0 mmol/L Mg2+、0.5 μmoL/L random primer、2.0 μL 10×PCR Buffer. The RAPD reaction was programmed by 1 cycle for 5 min at 94℃,followed by 45cycles for 30 s at 94℃,30 s at 37℃, and 70 s at 72℃and finally, followed by 1 cycle for 10 min at 72℃. The reaction system was reproducible and highly reliable, and it could be effectively for RAPD analysis in Taraxacum kok-saghyz Rodin.