亚麻SRAP反应体系的优化

Optimization of SRAP System in Flax

通过研究亚麻SRAP反应体系中主要因子对扩增结果的影响,建立了亚麻SRAP-PCR反应的优化体系。在20μL的反应体系中将PCR的5个主要成分分别设定8个浓度梯度,结果表明,最适宜的优化浓度分别为:1.5mmol/LMg2+、0.3mmol/LdNTP、1.5UTaq酶、30ng/μL模板DNA90ng和25ng/μL引物100ng。用6个亚麻材料验证优化体系,检测结果显示,多态性高,反应体系的稳定性和可重复性好,为SRAP标记技术在亚麻分子生物学研究方面的应用奠定了基础。

Studying on effect of the major elements of SRAP reaction system in flax, and establish an optimum SRAP-PCR system. Designing eight different concentrations respectively for five main factors of PCR in 20 μL reaction system, the results showed that the best concentration was 1.5 mmol/L Mg^2＋, 0.3 mmol/L dNTP, 1.5 U Taq enzyme, 30 ng/μL template DNA 90 ng, 25 ng/μL primer 100 ng. Using six flax germplasm to test this optimized SRAP system, the results showed that not only has the high polymorphism but also steady and reproducible. This work is a foundation for applying SRAP marker to sesame molecular biology studies.