蜡梅SSR反应体系建立及引物筛选

摘要

以素心蜡梅为材料,分别通过对影响蜡梅SSR扩增效果的模板DNA,dNTP,引物浓度,Taq酶和退火温度等因素的筛选,建立了其适宜的蜡梅SSR分子标记反应体系,同时筛选了适宜蜡梅SSR扩增引物.结果表明,蜡梅SSR扩增的适宜退火温度为55 ℃,在20μL反应体系中,蜡梅模板DNA质量浓度、dNTP引物浓度分别为5mg·L-1,0.2 mmo·L-1和0.5 μmol·L-1,Taq酶和10×Taq酶缓冲液量则为2 U和2.5μL.利用建立的反应体系,从不同木本植物SSR扩增的417对引物中筛选出了适合蜡梅SSR扩增的41对引物.%A study is made using Chimonanthus praecox as the materials, through the selection of template DNA, dNTP, primer concentration, Taq enzyme and annealing temperature for the impact of SSR amplification, establishing appropriate molecular markers reaction system of Chimonanthus SSR, and screening suitable SSR primers of Chimonanthus. The result showed that the appropriate annealing temperature of Chimonanthus SSR amplification was 55 ℃. In 20 μL reaction system, Chimonanthus template DNA, dNTP, primer concentration was 5 mg·L-1 ,0.2 mmol·L-1 and 0.5 μmol·L-1 ,Taq enzyme and 10 × Taq enzyme buffer was 2 U and 2.5 μL. Using the reaction system, 41 pairs of SSR primers for Chimonanthus SSR amplified were screened from 417 pairs different woody plants primers.