杨树RAMP体系的建立与优化  

Optimization of RAMP Reaction System in Poplar

作　　者：侯万伟[1] 樊军锋[1] 张小娟[2] 李雪林 

机构地区：[1]西北农林科技大学林学院,陕西杨陵712100 [2]西北农林科技大学植保学院,陕西杨凌712100 [3]青海省湟源县林业站,青海湟源812100

出　　处：《华北农学报》2008年第B10期217-219,共3页Acta Agriculturae Boreali-Sinica

基　　金：国家"十一五"科技支撑子专题(2006BAD08A11103)

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。Using poplar genome DNA as template,the major components of RAMP,such as concentrations of Mg2＋,dNTPs,TaqDNA polymerase,primers,were optimized in this study by orthogonal design in four factors three levels respectively.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 μmol/L in the 25 μL volume reaction.The amplification protocol was：94℃ 5 min,45 cycles for 1 min at 94℃,1 min at 45℃,2 min at 72℃,followed by a final extension of 10 min at 72℃.

关 键 词：杨树 RAMP 优化 

分 类 号：S792.11[农业科学—林木遗传育种]