二次通用旋转组合设计优化辣椒SRAP-PCR反应体系

采用二次通用旋转组合设计,对辣椒SRAP反应体系中的Taq酶用量、Mg2+浓度、dNTPs浓度、引物浓度和模板DNA用量等5个主要因素进行优化。结果表明,筛选出的辣椒最优SRAP-PCR反应体系为Taq聚合酶0.5 U,Mg2+2.3 mmol.L-1,dNTPs 0.2 mmol.L-1,引物0.8μmol.L-1,模板DNA 45 ng,总体积10μL。应用该体系对辣椒18份种质材料进行验证,结果证明该体系可靠稳定。因此,该试验的优化方法和优化体系适用于辣椒SRAP分子标记的研究。

Optimization of SRAP-PCR system in pepper by quadratic general rotary unitized design

The quadratic general rotary unitized design was used to optimize SRAP-PCR system in pepper(Capsicum spp.) with five key factors(Taq DNA polymerase,Mg2+,dNTPs,primer and template DNA,respec-tively).The results showed that the optimized SRAP-PCR system for pepper was: 0.5 U Taq DNA polymerase,2.3 mmol?L-1 Mg2+,0.2 mmol?L-1 dNTPs,0.8 μmol?L-1 primer,45 ng template DNA in a total of 10 μL reaction solu-tion.The optimized SRAP-PCR system was tested on 18 pepper germplasm and was reliable and steady.Accord-ingly,the method of the test and the optimized SRAP-PCR system were suitable for SRAP(sequence-related am-plified polymorphism) analysis of pepper.