All factors affecting the PCR system in Pinus massoniana Lamb were investigated one by one.The results show that the optimum PCR system of EST-SSR were:2 μL 10×Buffer(Mg2+ Free),30 ng template DNA,0.187 5 mmol/L dNTPs,3.75 mmol/ L Mg2+,8 pmol primer pair,1.0 U Taq DNA poly-merase in total 20 μL reaction system.Finally,a total of 11 isolates of P.massoniana was used for testing the stability of the PCR amplification.The results prove that the optimum PCR system was stable,reliable,highly repetitive.
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