月季SSR反应体系的优化

Optimization of SSR Analysis System in China Rose

以月季品种‘月月粉’为试材,利用CTAB法提取DNA,对影响SSR-PCR扩增体系的主要因子设计了多梯度的优化实验,建立了适用于月季的SSR反应体系:总反应体积为25μL,包括模板DNA溶液(20 ng/μL)2μL,10×PCR buffer(Mg2+free)2.5μL,MgCl2(25 mmol/L)2.0μL,dNTPs(10 mM)0.5μL,正反引物各(10μmol/L)1μL,TaqDNA聚合酶(5 U/μL)0.3μL.利用该优化体系对部分月季品种进行PCR扩增和电泳检测,扩增结果清晰且有较高的多态性,表明该体系适合用于分析月季的遗传多样性.

DNA of Rosa ＇Pallida＇ was extracted by fication system were included in a multi-gradient suitable for China rose was established. The best CTAB. The main factors affecting the SSR-PCR amplioptimization experiment, and an SSR reaction system reaction system of SSR-PCR was. 2μL template DNA （20 ng/μL）, 2.5 μL 10×PCR buffer （Mg^2＋ free）, 2.0 ×L MgCl2 （25mmol/L）, 0.5 μL dNTPs （10 mM）, 1 μL each primer （10 μmol/L） and 1.5 U Taq polymerase in 25 μL reaction system. Using the above PCR system, SSR fragments of 12 China rose cultivars were obtained. The clear and abundant polymorphism indicated that this system was suitable for the analysis of genetic diversity in China rose.