回归分析在辣椒品种RAPD反应体系优化中的应用

Optimization of Applying Regressive Analysis to RAPD Reaction System in Studying Cultivars of Capsicum annuum

以辣椒(CapsicumannuumL.)为材料,采用回归分析的方法对RAPD体系进行优化研究。结果表明在体积为20μl反应体系中,主要因子的优化组合是:Mg2+、dNTP、Taq酶、引物、模板DNA等的浓度分别为3mmol/L、0.3mmol/L、1.3U、0.4μmol/L和64ng;热循环程序为:94℃预变性2min,94℃变性30s,40℃退火1min,72℃延伸90s,47个循环,最后72℃延伸10min。通过对24个辣椒品种进行遗传多样性分析,证明该优化体系是稳定可靠的。

Optimized RAPD system with the aid of regressive was used in studying cultivars of Capsicum annuum L. The results indicated that concentrations of Mg^2＋ , dNTP, Taq polymerase, primer, template DNA were 3 mmo/L, 0.3 mmol/L, 1.3 U, 0.4μmol/L, 64 ng respectively in 20μl reaction system and thermal cycle program was 1 cycle of denaturing for 2 min at 94℃, 47 cycles of denaturing for 30 s at 94℃, annealing for 1 min at 40℃, primer extension for 90 s at 72℃ and the last cycle of extension for 10 min at 72℃. The RAPD reaction system identified by genetic diversity analysis of 24 Capsicum cultivars was stable and dependable.