大白菜SRAP反应体系的建立与优化  

Optimization of SRAP Reaction System in Chinese Cabbage(Brassica campestris syn. rapa L. ssp. Pekinensis(Lour) olsson)

作　　者：杨琦[1] 张鲁刚[1] 

机构地区：[1]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《西北农业学报》2007年第3期119-123,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金资助项目(编号30270913)

摘　　要：以大白菜基因组DNA为模板,通过正交试验设计,从Mg2+、Taq酶、dNTPs、引物、模板5种因素4个水平对大白菜SRAP(Sequence-related amplified polymorphism,序列相关扩增多态性)反应体系进行优化,建立了适合于大白菜的SRAP-PCR优化反应体系,该体系为25μL:Mg2+浓度为3.0 mmol/L,dNTPs为0.2mmol/L,Taq酶1.5 U,引物为0.2μmol/L,模板DNA73.2 ng。PCR反应程序为:94℃预变性5 min;94℃变性1 min,35℃复性1 min,72℃延伸1 min,5个循环;94℃变性1 min,50℃复性1 min,72℃延伸1 min,35个循环,72℃延伸10 min。Using Chinese Cabbage （Brassica campestris syn. rapa L. ssp. Pekinensis （Lour） olsson） genome DNA as template, the major components of SRAP, such as concentrations of Mg^2＋ , dNTPs, Taq DNA polymerase, primers and template, were optimized in this study by orthogonal design in five factors four levels respectively. The results showed that the optimum SRAP reaction system includes Mg^2＋ 3.0 mmol/L, dNTPs 0.2 mmol/L, DNA template 73.2 ng,Taq DNA polymerase 1.5 U and primer 0.2 μmol/L in the 25μL volume reaction. The most suitable protocol was initially denaturing at 94℃ for 5 min, then pre-amplifying at 94℃ 1 min, 35℃ lmin and 72℃ 1 min for five cycles, finally amplifying for 35 cycles when the annealing temperature was adjusted to 50℃.

关 键 词：大白菜 SRAP 优化 正交试验 

分 类 号：S634.1[农业科学—蔬菜学]