摘要： 以萝卜基因组DNA为模板,对其SRAP-PCR反应体系的各个主要影响因子进行了系统的优化,建立了重复性好、多态性丰富的萝卜SRAP-PCR反应体系:20μL反应体系中,DNA50 ng、Mg²2.25 mmol/L、dNTP 0.2 mmol/L、Taq1 U、Primer 0.7μmol/L。扩增程序为:94℃预变性5 min;94℃变性1 min、35℃复性1 min、72℃延伸1 min,5个循环;94℃变性1 min、50℃复性1 min、72℃延伸1 min,35个循环,72℃延伸10 min,4℃保温。该反应体系在不同萝卜种质资源遗传多样性分析、遗传图谱构建、分子标记辅助选择育种等方面都有重要作用。

关键词: 萝卜, SRAP-PCR, 反应体系

Abstract: In this study,radish genomic DNA was used for the PCR template to optimize radish SRAP -PCR reaction system.A SRAP -PCR system with good stability was established.In the 20 μL reaction system ,DNA 50 ng ,Mg2+2.25mmol/L,dNTP 0.2 mmol/L,Taq 1 U,Primer 0.7 μmol/L.PCR procedure :initially denaturing at 94 ℃ for 5 min ;then94 ℃, 1 min,35 ℃, 1 min and 72 ℃, 1 min for the first five cycles,then the annealing temperature was raised to 50 ℃ for another 35 cycles;At last,72 ℃10min and 4 ℃for ever.The optimum SRAP -PCR reaction system is adaptive to different radish variety.It laid a good foundation for genetic diversity analysis,genetics map construction and molecular assisted breeding in radish.

Key words: Radish(RaphanussativusL.), SRAP-PCR, Reactionsystem

中图分类号: 

• S631.03