摘要
以美国红枫基因组DNA为模板,采取L16(45)正交实验设计,对影响扩增反应的5个主要因素进行优化。结果表明:在20μL反应体系中,各因素的最佳配比为Mg2+浓度1.5mmol/L,dNTPs浓度2.5mmol/L,引物浓度1.5μmol/L,模板DNA用量50ng,Taq DNA聚合酶用量0.5U。该体系的建立为SRAP技术在美国红枫分子辅助育种实践中提供了技术基础。
Using the genomic DNA of Acer rubrum as template, L16 （4^5） orthogonal experiment was designed to optimized the 5 major factors influencing the amplification reaction. The results showed that the optimum ratio of each factor in 20 μL reaction system was 1.5 mmol/L of Mg^2＋ concentration,2. 5 mmol/L of dNTPs concentration,1. 5 μmol/L of Primer concentration,50 ng of template DNA,and 0. 5 U of Taq DNA polymerase. The establishment of this system provided a technological basis for the application of SRAP in molecular assisted breeding in Acer rubrum.
出处
《北方园艺》
CAS
北大核心
2015年第2期79-83,共5页
Northern Horticulture
基金
国家林业局"948"资助项目(2011-4-25)
辽宁省科技计划资助项目(201404214)
