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[目的]建立一套适合甘草分子学研究的RAPD-PCR反应体系。[方法]以甘草种质为试材,采用正交试验法设计,对影响RAPD-PCR扩增的主要因素dNTPs、引物、Taq酶和DNA模板进行优化筛选。[结果]总体积25μl的甘草RAPD-PCR最佳反应体系为:10×PCR缓冲液(含MgCl2)2.5μl,10mmol/LdNTPs2.5μl,50ng DNA2μl,10μmol/L引物2μl,5UTaq酶0.4μl。对引物的退火温度进行了梯度筛选,34℃时扩增效果较好。[结论]进行甘草RAPD-PCR反应体系的正交优化非常有效。
[Objective] To establish a RAPD-PCR reaction system suitable for the molecular study of licorice. [Method] Glycyrrhiza uralensis germplasm was used as test material, and the orthogonal test was used to optimize screening of dNTPs, primers, Taq enzyme and DNA template that affect RAPD-PCR amplification. [Result] The optimum reaction system of RAPD-PCR for licorice root in 25μl total volume was 2.5μl of 10 × PCR buffer (containing MgCl2), 2.5μl of 10mmol / L dNTPs, 2μl of 50ng DNA, 2μl of 10μmol / L primer and 0.4μl of 5UTaq enzyme. The primer annealing temperature gradient screening, amplification effect is better at 34 ℃. [Conclusion] The orthogonal optimization of RAPD-PCR reaction system was very effective.