枣树ISSR反应体系的建立及优化

Establishment and Optimization of an ISSR Reaction System for Chinese Date(Ziziphus jujuba) Cultivars

为建立枣树适宜的ISSR反应体系,以枣树叶片为材料,对ISSR反应体系中的主要影响因子进行了优化。研究了引物浓度、dNTP浓度、TaqDNA聚合酶及退火温度对扩增的影响。结果表明,在25μLISSR体系中各组分的适宜的浓度为:1×PCR buffer,375μmol/LdNTP,0.2μmol/L引物,1.5 UTaqDNA聚合酶。不同引物间适宜的退火温度不同。应用该ISSR体系对10个不同品种的枣材料进行了扩增,证实了该体系的适用性和稳定性。

In this study,ISSR conditions on Chinese date were optimization.The main effectors such as the concentration of primer,dNTP,Taq DNA polymerase were screened to optimize ISSR amplification system by orthogonal design experiments as materials.The annealing temperatures were investigated. The results showed that the optimum concentrations of components in 25 μL ISSR reaction system were as follows：1×PCR buffer,375 μmol/L dNTP,0.2 μmol/L primer,1.5 U Taq DNA.The annealing temperatures varied in different primers.The system developed was successfully applied in the amplification of 10 cultivars of chinese date,indicating the suibtability of the system.