枸杞SRAP反应体系建立和优化

以宁杞1号为试验材料,探讨Mg2+、dNTPs、Taq DNA聚合酶用量、引物浓度、模板DNA用量对枸杞SRAP-PCR反应的影响.建立20μL反应体系,模版DNA 50 ng,Buffer 1×,Mg2+1.5 mmol.L-1,dNTPs 0.3 mmol.L-1,引物0.3μmol.L-1,Taq DNA聚合酶用量为1 U,并利用该反应体系,两对不同引物组合Me1/Em1和Me3/Em5对12份枸杞样品DNA进行SRAP-PCR扩增,1.8%琼脂糖凝胶电泳检测结果表明,不同品种(系)间DNA谱带多态性丰富,说明该体系稳定可靠.

Establishment and optimization of SRAP-PCR reaction system in Chinese wolfberry

Taking Ning Qi 1 as material,the concentrations of Mg2+,dNTPs,Taq DNA polymerase,primers,template DNA which affected the SRAP-PCR reaction were optimized in order to establish the SRAP molecular system in Chinese wolfberry(Lycium bararum L).The optimum system was as follows(20 μL): template DNA 50 ng,Buffer 1×,Mg2+ 1.5 mmol·L-1,dNTPs 0.3 mmol·L-1,primer 0.3 μmol·L-1,Taq DNA polymerase 1 U.Through above SRAP-PCR system,primer combinations Me1/Em1 and Me3/Em5 were used to amplify 12 Chinese wolfberry cultivars.Polymorphism between different cultivars was abundantly detected by 1.8% agarose gel.The results indicated that this system was stable and suitable.