辽东栎SSR反应程序和反应体系的优化

The Optimization of the SSR Reaction Program and the Reaction System in Researches on Quercus liaotungensis

本文针对辽东栎遗传多样性研究过程中的SSR反应程序和反应体系进行了优化研究。优化后的反应程序为:94℃预变性3 min;94℃变性45s,58℃退火45 s,72℃延伸7 min,72℃延伸45 s,33个循环;经优化后的反应体系为30 ngDNA模板,2μL10×Buffer(Mg2+Plus),150μmol·L-1dNTP Mixture,0.3μmol·L-1上下游引物,1U(5u·μL-1)Taq酶,反应总体积为20μL。研究结果对SSR分子标记在栎属植物遗传多样性中的应用也有一定的参考价值。

In this paper,optimization researches were conducted on the SSR reaction program and the reaction system for the research process of Q. liaotungensis genetic diversity. The optimized reaction program was to adopt 33 cycles,which included 3 minutes of pre-denaturing at 94℃ ,45 seconds of denaturing at 94℃ ,45 seconds of annealing at 58℃ ,45 seconds of extension at 72℃, and a final extension step was carried out at 72℃ for 7 minutes. The optimized reaction system was as follows：30 ng DNA,2 μL10 × Buffer （ Mg^2＋ Plus）, 150 mol · L ^- 1 dNTP Mixture, 0. 3 μmol · L^ -1 of each primer, 1U （ 5 μ·μL-1 ） Taq polymerase, and the total reaction volume was 20 p.L. The results had a certain reference value for the ap- plication of the SSR marker in the research on the genetic diversity of oak plants.