山核桃SSR反应体系优化

Optimization on SSR Reaction System of Carya cathayensis

优化SSR反应体系是山核桃SSR遗传图谱构建的基础.通过对PCR反应中Taq聚合酶用量、dNTP浓度、引物浓度、Mg2+浓度和模板DNA量的组合试验,确定了山核桃SSR的最佳反应体系,即在30μL反应体系中,含600 ng模板DNA,1×PCR Buffer,2.0UTaq聚合酶,0.22mmol/L dNTP,1.0 mmol/L Mg2+,0.13μmol/L引物.

Optimization on SSR reaction system is the fundamental step in SSR genetic map construction for Carya cathayensis. The systematic experiments on dosage of Taq DNA polymerase, dNTP concentration, concentrations of the primer, Mg^2＋ and template DNA and other factors were carried out to screen the optimal reaction system. It was indicated by the results that the optimal reaction system of SSR for Carya cathayensis was 30μL of total reaction volume containing 600 ng of template DNA, 1× PCR Buffer, 2. 0 units of Taq DNA polymerase, 0. 22 mmol/L of dNTP, 1.0 mmol/L of Mg^2＋ and 0. 13 μmol/L of primer.