东北山葡萄SSR反应体系的建立及优化

以东北山葡萄(左山二)叶片为材料,对SSR反应体系中的主要影响因子进行了优化。研究了模板浓度、引物浓度、dNTP浓度、TaqDNA聚合酶量对扩增的影响。结果表明,在20μL SSR体系中各组分的适宜浓度为:1×PCR buffer,引物0.3μmol/L,模板DNA 30 ng,dNTP 0.2 mmol/L,TaqDNA聚合酶1.0U。应用该SSR体系,用3对引物对5份山葡萄材料进行了扩增,证实了该体系的适用性和稳定性。

Establishment and Optimization of an SSR Reaction System for Vitis amurensis

The main factors in SSR on Vitis amurensis were optimized using the leaves of grape eultivar'Zuoshaner'as material in this study.The concentrations of primer,DNA,dNTP and the dosage of TaqDNA polymerase were screened to optimize SSR amplification system.The results showed that the optimum concentrations of components in 20uL SSR reaction system were as follows:1 x PCR buffer,0.3umol/L primer,30ng DNA,0.2mmol/L dNTP,1.0U TaqDNA polymerase.The system developed was successfully applied in the amplification of 5 cuhivars of Vitis amurensis with three pairs of primer.This indicated the suitability and steadiness of the system.