思茅松SSR-PCR反应体系优化研究

Optimization of SSR-PCR Reaction System in Pinus kesiya var. langbianensis

以思茅松杂交亲本JG1号基因组DNA为模板,利用正交设计实验对影响SSR-PCR反应体系的主要因子进行了优化,建立了一套最佳思茅松SSR-PCR反应体系。结果表明:在25μL最佳反应体系中,各组分最佳含量分别为Mg^(2+) 2.0 mmol/L、DNA 60 ng、引物0.50μmol/L、Taq DNA聚合酶0.25U、dNTP 0.20 mmol/L。利用11份个体对此SSR反应体系进行验证,获得扩增谱带清晰且多态性较好,证明该体系稳定可靠。

By using the genomic DNA of JG1 as template,an orthogonal design was employed to optimize SSR-PCR system of Pinus kesiya var.langbianensis,and the optimum SSR-PCR reaction system of P.kesiya var.langbianensis was established.The results showed that the optimum reaction system was 25^L reaction solution containing Mg2+2.0 mmoL^L,DNA 60 ng,primer 0.50^moL/L,Taq DNA polymerase 0.25 U,dNTP 0.20 mmoL^L.To verify the reaction system DNA of 11 individuals were amplified,the results indicated that the optimized reaction system was proved in good stability.