甜叶菊RAPD反应体系的优化

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, 20 μL的优化体系包括: 双蒸水13.6 μL, 10×Buffer(含15 mmol/L MgCl2)溶液3 μL, 2.5 mmol/L的dNTPS 1.2 μL, 10 μmol/L的引物1 μL, 20 ng的模板DNA 1 μL, 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 ℃延伸1 min, 40个循环, 最后72 ℃延伸7 min。

Abstract

Optimization of 7 factors of RAPD reaction system in Stevia rebaudiana Bertoni was studied. The results indicated that, optimization system of 20 μL included: 13.6 μL of ddH20, 3 μL of 10×Buffer with 15 mmol/L MgCl2, 1.2 μL of 2.5 mmol/L dNTPS, 1 μL of 10 μmol/L Primer, 1 μL of 20 ng DNA and 1 unit of Taq polymerase; thermal cycle program was: 1 cycle of denaturing for 4 min at 94 ℃, 40 cycles of denaturing for 30 s at 94 ℃, annealing for 30 s at 35 ℃, primer extension for 1 min at 72 ℃, and the last cycle of extension for 7 min at 72 ℃.

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胡能兵, 何克勤, 张子学, 林平, 隋益虎, 崔广荣. 甜叶菊RAPD反应体系的优化[J]. 激光生物学报, 2011, 20(4): 560. HU Nengbing, HE Keqin, ZHANG Zixue, LIN Ping, SUI Yihu, CUI Guangrong. Optimization of RAPD Reaction System in Stevia rebaudiana Bertoni[J]. Acta Laser Biology Sinica, 2011, 20(4): 560.

甜叶菊RAPD反应体系的优化

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