摘要： 以杨树基因组DNA为模板,通过正交试验设计,从Mg2+、Taq酶、dNTPs、引物4种因素3个水平对杨树RAMP反应体系进行优化,建立了适合于杨树的RAMP-PCR优化反应体系,该体系为25μL:Mg2+浓度为1.5 mmol/L,dNTPs为0.2 mmol/L,Taq酶1.0 U,引物各为0.4μmol/L。PCR反应程序为:94℃预变性5 min,94℃变性1 min,45℃复性1 min,72℃延伸2 min,45个循环,72℃延伸10 min。

关键词: 杨树, RAMP, 优化

Abstract: Using poplar genome DNA as template, the major components of RAMP, such as concentrations of Mg²⁺,dNTPs, TaqDNA polymerase, primers, were opt imized in this study by orthogonal design in four factors three levels re??spectively.The results showed that the optimum RAMP reaction system includes Mg²⁺ 1.5 mmol/ L, dNTPs 0.2 mmol/ L,TaqDNA polymerase 1.0 U and primer 0.4 ??mol/L in the 25L volume reaction.The amplification protocol was: 94.5min, 45 cycles for 1 min at 94.1 min at 45.

Key words: Poplar, RAMP, Optimization

中图分类号: 

• S792.11