油葵SSR-PCR反应体系的优化及引物筛选

Optimization of SSR-PCR Reaction System and Primer Screening in Oil Sunflower

为了获得适合油葵的SSR-PCR反应体系,通过采用控制变量法对影响SSR反应体系的主要因素进行优化,研究Mg2+浓度、d NTPs浓度、Taq酶浓度、引物浓度、退火温度等因素。结果表明各因素的终浓度分别在Mg2+浓度2.5 mmol/L、d NTPs浓度0.2 mmol/L、Taq酶量0.5 U、引物浓度0.4μmol/L及退火温度为53.9℃时最为适合。通过试验优化后的体系筛选引物,最终从50对引物筛得19对多态性好的引物可进行下一步的研究。

In order to obtain the SSR-PCR reaction system for oil sunflower, the method of control variable hadbeen used to optimize the main factors influencing the SSR reaction system, which include the concentration of Mg^2＋, d NTPs, Taq enzyme, primer, and annealing temperature. Results showed that the optimized SSR reactionsystem was Mg2＋2.5 mmol/L, d NTPs 0.2 mmol/L, Taq polymerase 0.5 U, primer 0.4 μmol/L and the annealingtemperature 53.9℃. Through the testing system after optimization, 19 pairs of polymorphic good primer couldbe used for the next research from 50 pairs of primers screening.