甜叶菊RAPD反应体系的优化

Optimization of'RAPD Reaction System in Stevia rebaudiana Bertoni

以甜叶菊为试材,对影响其RAPD反应体系的7个因子进行优化。结果表明,20μL的优化体系包括:双蒸水13.6μL,10×Buffer(含15 mmol/L MgCl_2)溶液3μL,2.5 mmol/L的dNTPS 1.2μL,10μmol/L的引物1μL,20 ng的模板DNA 1μL,1 U Taq聚合酶;热循环程序为:94℃预变性4 min,94℃变性30 s,35℃退火30 s,72℃C延伸1 min,40个循环,最后72℃延伸7 min。

Optimization of 7 factors of RAPD reaction system in Stevia rebaudiana Bertoni was studied. The results in- dicated that, optimization system of 20 ~xL included：l 3.6 /μL of ddH20, 3 μL of 10 x Buffer with 15 mmol/L MgC12, 1.2 μL of 2.5 mmol/L dNTPS, 1 μL of 10 p^mol/L Primer, 1 μL of 20 ng DNA and 1 unit ofTaq polymerase; thermal cycle program was ： 1 cycle of denaturing for 4 min at 94 °C, 40 cycles of denaturing for 30 s at 94 °C, annealing for 30 s at 35 °C, primer extension for 1 min at 72°C, and the last cycle of extension for 7 min at 72 °C.