芒果ISSR反应体系的建立与优化

Optimization of ISSR Reaction System for Mango

运用L16(45)正交设计对芒果ISSR反应的5个因素,即DNA模板浓度、Mg2+浓度、dNTPs浓度、引物浓度、Taq DNA聚合酶浓度在4个水平上进行优化试验,通过不同反应体系扩增效果比较,建立优化的芒果ISSR反应体系,即25μL的PCR体系中含有DNA模板25 ng、Mg2+浓度为1.5 mmol/L、dNTPs 0.3 mmol/L、引物0.32μmol/L、Taq DNA聚合酶1.25 U。这为进一步运用ISSR标记在DNA分子水平上对芒果种质资源进行分析奠定基础。

The orthogonal design of five factors in four levels was used to optimize the ISSR-PCR amplification system on mango.The factors were the concentrations of DNA template,Mg2＋,dNTPs,primer and TaqDNA polymerase.The results showed that： the optimal concentration of the five important components was 25 ng,1.5 mmol/L,0.3 mmol/L,0.32 μmol/L,1.25 U,respectively in a 25 μL ISSR reaction system.This research laid a foundation for mango germplasm identification and analysis at DNA level.