高山红景天RAPD反应体系的优化研究

Study on RAPD Reaction System for Endangered Pharmacy Plant Rhodiola Sachalinensis

[目的]筛选适合高山红景天的RAPD-PCR反应体系。[方法]以药用植物高山红景天雌株的叶片为材料,采用改良的CTAB法提取基因组DNA,并对影响RAPD扩增反应的各因素进行优化。[结果]结果表明,适合高山红景天的RAPD反应体系为:25μl总体积中含DNA模板20 ng,Mg2+1.5 mmol/L,引物10 pmol,dNTP 100μmol/L,Taq DNA聚合酶1.0 U。[结论]CTAB法在高山红景天可获得质量较好的基因组DNA。

[Objective] The study aimed to choose RAPD-PC reaction system of fit Rhodiola sachalinensis. [Method]With the leaves of genomic plant Rhodiola sachalinensis as materials,the genomic DNA was extracted from young leaves of endangered medicinal plant Rhodiola sachalinensis by means of improved CTAB method. The RAPD reaction system for Rhodiola sachalinensis was optimized including cycling program and the concentration of template, primer, dNTP, Taq DNA polymerase and Mg^2＋ . [ Result ]The results indicated that the RAPD reaction system were 20 ng of DNA template, 1.5 mmol/L of Mg^2＋ ,primer 10 pmol,dNTP 100μmol/L,Taq DNA polymerase 1.0 U.[Conclusion] The CTAB will obtain high quality genome team DNA in Rhodiala sachalinensis.