摘要
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。
The quadratic general rotary unitized design was used to optimize SRAP-PCR system in pepper （Capsicum spp.） with five key factors （Taq DNA polymerase, Mg^2＋, 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 Mg^2＋, 0.2 mmol.L^-1 dNTPs, 0.8μmol.L^-1 primer, 45 ng template DNA in a total of 10 μL reaction solution. 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.
出处
《安徽农业大学学报》
CAS
CSCD
北大核心
2012年第3期478-482,共5页
Journal of Anhui Agricultural University
关键词
辣椒
SRAP
二次通用旋转组合设计
体系优化
pepper
SRAP
quadratic general rotary unitized design
system optimization
