白梭梭ISSR-PCR实验反应体系的建立和优化

Establishment and Optimum of PCR Reaction System in Haloxylou persicum

探讨了白梭梭ISSR实验中的各种影响因素,采用正交设计L16(45)对PCR反应中5个因素(Taq酶,Mg2+,模板DNA,dNTP和引物)在4个水平上进行优化试验。在初步实验的基础上通过因素内比较进一步优化各因素在ISSR-PCR反应中的浓度;设置了不同的退火温度和循环次数,建立了白梭梭ISSR的最佳反应体系,为进一步进行白梭梭种群间遗传多样性的研究奠定基础。

This article focuses on the influential factors of ISSR experiments used to analyze Haloxylou persicum. The orthogonal design was used to optimize ISSR-PCR amplification system in four levels of five factors （Taq DNA polymerase, Mg^2＋ , DNA template, dNTP and primer） respectively. Based on the preliminary experiment,the density within factors of ISSR-PCR was further optimized. And different annealing temperature and cycle times were also compared. The reaction condition of ISSR experiment was discussed, which was basis for the study on genetic diversity of Haloxylou persicum populations.