思茅松SSR-PCR反应体系优化研究

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

摘要: 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25 μL最佳反应体系中，各组分最佳含量分别为Mg²⁺ 2.0 mmol/L、DNA 60 ng、引物0.50 μmol/L、Taq DNA聚合酶0.25 U、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。

Abstract: 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 μL reaction solution containing Mg²⁺ 2.0 mmol/L, DNA 60 ng, primer 0.50 μmol/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.