Extraction of DNA and optimization of RAPD reaction system for Phellinus igniarius

Mycelium of Phellinus igniarius was used as experimental material,the CTAB and SDS methods was used to extract DNA,and the reaction system of RAPD was optimized.The results showed that the extracted genomic DNA was pure and integral in the CTAB method,and the optimum reaction system was established as follows(25μL reaction volume):1.0μL template DNA,0.8μL primer,0.6μL dNTPs,0.3μL Taq DNA polymerase,2.5μL Buffer and 19.8μL ddH2O.Orthogonal experiment was used to validate the stability and currency of this system.The PCR program was 5min at 94℃ for predenaturalization,then 45 cycles of 1min at 94℃ for denaturation,1min at 40℃ for annealing and 1.5min at 72℃ for extension,finally extension at 72℃ for 5min.