余甘子SRAP反应体系的优化

Optimization of Sequence-Related Amplified Polymorphism Amplification System for Phyllanthus emblica

为建立余甘子SRAP-PCR反应的最优体系,通过正交设计和单因素试验调整优化PCR反应各因素(模板DNA、Mg^(2+)、dNTPs、引物)。结果表明,各因素对扩增结果均有不同的影响。采用50ng模板DNA,2.0mmol/L Mg^(2+),0.4 mmol/L dNTPs,0.2 μmol/L引物的20μL反应体系可扩增出最清晰丰富的多样性条带。使用该优化体系检测12份余甘子种质资源样品,结果稳定可信,适用于分子遗传学研究。

The optimized SRAP-PCR amplification system for Phyllanthus emblica was established by the orthogonal design and single factor optimization of PCR factor（template DNA,Mg^2 ＋,dNTPs,primers）.It revealed that each PCR factor had different effect on the PCR results,and 50 ng template DNA,2.0 mmol/L Mg^ 2 ＋,0.4 mmol/L dNTPs,0.2 μ mol/L primers in a total of 20 μ L reaction solution amplified the most rich polymorphism and clear bands.The optimized SRAP-PCR system was tested on twelve Phyllanthus emblica germplasms and proved to be steady and reliable for molecular genetics research.