油葵SSR反应体系的优化

采用改良CTAB法从油葵种仁中提取基因组DNA,用于SSR-PCR反应。对影响油葵SSR-PCR反应的几个重要因子进行了探讨,确定了最佳的反应体系:在总体积为25μL中,DNA为2.0 ng/μL,Mg2+浓度为1.5 mmol/L,dNTPs浓度为0.2 mmol/L,Taq酶量为2.0 U,引物浓度为0.25μmol/L,退火温度为52～54℃,扩增的谱带清晰、多态性丰富。该SSR-PCR反应体系为油葵亲缘关系分析奠定了技术基础。

Optimization of SSR analysis system in oil sunflower

In this study,the top-quality genomic DNA was extracted from seeds of oil sunflower with an improved method of CTAB,and the SSR analysis system was optimized.The factors,which vitally affected the SSR results were studied.And the optimal results are as follows: DNA 2.0 ng/μL,Mg2+ 1.5 mmol/L,dNTPs 0.2 mmol/L,Taq polymerase 2.0 U and primers 0.25 μmol/L in reaction mixture of 25 μL and annealing temperature 52~54 ℃.Using the above PCR system,SSR pattern was clear and abundant.Thus,the optimal SSR analysis system provided some technical foundations to study the phylogenetic relationship of oil sunflower.