八仙花SRAP反应体系的建立与优化

Optimization of SRAP Reaction System in Hydrangea

为建立适合八仙花SRAP-PCR分子标记技术体系,以八仙花栽培品种H.macrophylla‘Lavbla’为材料,通过单因子实验分别研究了模板DNA、dNTPs、Mg2+、Taq酶浓度和引物用量对八仙花SRAP扩增反应的影响,确立了八仙花SRAP分析的最佳反应体系为25μl:模板DNA60ng、Mg2+2.0mmol/L、dNTPs0.7mmol/L、Taq酶0.5U、引物0.7μmol/L×2、10×PCRBuffer2.5μl,该反应条件下八仙花SRAP扩增条带清晰,多态性丰富。

Taking hydrangea cultivated variety H.macrophylla ＂Lavbla＂ as the material, using the single factor experiment to study influence that template DNA. dNTPs, Mg^2＋, Taq enzyme concentration and the dosage of primers respectively to the hydrangea SRAP reaction. SRAP reaction system in hydrangea was established as follow： template DNA 60 ng, Mg^2＋ 2.0 mool/L, dNTPs 0.7 mool/L, Taq enzyme 0.5 U, primer 0.7 μmol/L, 10×PCRBuffer 2.5 μl, under this response condition the hydrangea SRAP amplified band was clear and the polymorphism was rich.