洋葱SRAP-PCR体系的优化研究

Optimization of SRAP-PCR System for Onion

优化了洋葱SRAP-PCR反应体系的模板浓度和Mg2+浓度。采用8对不同的引物组合和2份不同的洋葱材料进行验证实验,结果表明,在20μl的反应体系中,模板的量在180～210 ng均可获得清晰的扩增,低于180 ng,扩增带模糊,高于210 ng扩增背景严重;合适的Mg2+浓度在1.5～2.0 mmol/L之间;使用优化的反应体系,扩增条带清晰,可满足不同引物组合和洋葱材料的要求。

The concentrations of DNA-template and Mg2＋ were optimized to establish the SRAP molecular marker system in onion.The system had been verified by eight pairs of primers and two onion materials.In 20 μl reaction system,180～210 ng DNA template and 1.5～2.0 mmol/L Mg2＋ were more suitable for onion SRAP marker system.The amplified fragments obtained by this optimized system were clear,distinguishable,steady and reliable for different primer combinations and onion varieties.