DNA Extraction and Optimization of RAPD Reaction System for Phoebe zhennan
DNA of Phoebe zhennan was extracted from the fresh leaves and silica gel dried leaves,and the various factors affecting RAPD reaction was optimized.The optimized reaction system and programme of RAPD for Phoebe zhennana were as follows：each 25 μL amplification reaction solution suited for Phoebe zhennana RAPD was consisted of 2μL DNA,0.3 μLTaq DNA polymerase,1 μL primer,5 μL MgCl2,0.5 μL dNTPs and 16.2 μL ddH2O.The PCR amplification program suited for Phoebe zhennan RAPD were pre-denaturing at 94 ℃ for 3 min,followed by denaturing at 94 ℃ for 1 min,annealing 36 ℃ for 1 min,extending 72 ℃ for 2 min and cycling 42 times,and lastly,extending 72 ℃ for 10 min and storing at 4 ℃.The results showed that same results of PCR amplification could be got from the fresh leaves and silica gel dried leaves and could meet the need for researches.
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