湖北海棠SRAP-PCR反应体系的优化

Optimization of SRAP-PCR Reaction in Malus hupehensis

对PCR反应中相互影响的三因素采用三水平全组合设计,建立了湖北海棠SRAP-PCR的最佳反应体系:Mg2+2.5mmol·L-1,Taq酶1.5U,dNTP0.3mmol·L-1,引物0.3μmol·L-1,Buffer1×,DNA模板30ng,反应总体积25μL。应用该体系对33个湖北海棠样本进行扩增,得到了清晰、多态性丰富的扩增条带,证明该体系稳定可靠,可以在湖北海棠的相关研究中应用。

In this research, tile complete combination design of 3 factors with 3 leveles in PCR reaction was adopted and the best SRAP-PCR reaction system for Mdus hupehensis was established. That was Mg^2＋ 2.5 mmol·L^-1, Taq polymerase 1.5U, dNTP 0.3 mmol·L^-1, each primer 0.3 μmol·L^-1, Buffer lx, DNA template 30 ng, total reaction volume 25 μL. By amplifying the 33 M. hupehensis using this reaction system, clear and polymorphic amplification bands were obtained. The stability and reliability of the reaction system was confirmed.