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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.
In this study, we used the CTAB method to extract DNA from the cultivars “Moon Moon-pink” and designed the multi-gradient optimization experiments on the main factors influencing the SSR-PCR amplification system. The SSR reaction system suitable for rose was established. The total reaction volume (25 μL), 2.5 μL of 10 × PCR buffer (Mg2 + free), 2.0 μL of MgCl2 (25 mmol / L) and 0.5 μL of dNTPs (10 mM) 1 μL of Taq DNA polymerase (5 U / μL) and 0.3 μL of Taq DNA polymerase (5 U / μL) were used for PCR amplification and electrophoresis of some rose cultivars with clear and high amplification results, indicating that the system Suitable for analyzing the genetic diversity of roses.