冷暖型小麦基因差异的最佳RAPD反应体系研究  

Study on RAPD Reaction System for Gene Difference of the Cold and Warm Type of Wheat

作　　者：裴国亮[1] 李京敬[1] 王长发[1] 陶士珩[1] 

机构地区：[1]西北农林科技大学生物信息中心,陕西杨凌712100

出　　处：《安徽农业科学》2006年第13期2993-2995,共3页Journal of Anhui Agricultural Sciences

摘　　要：为了建立小麦RAPD反应的最佳体系,保证反应结果的稳定性和可靠性,寻找小麦冷暖型基因差异,降低试验成本,在相同PCR扩增程序(94℃预变性5 min,94℃变性45 s,38℃退火1 min,72℃延伸1.5 min,40个循环,72℃延伸10 min,4℃保存)下,对冷暖型小麦基因组DNA的RAPD扩增体系各参数进行比较筛选,创建其最佳反应体系为:25μl反应体系中,模板DNA 20 ng,10×PCR缓冲液2.5μl,dNTPs 0.2 mmol/L,Taq酶1 U,Mg2+浓度为2.0 mmol/L,ddH2O 12μl,随机引物10 ng。In this research, with the same PCR program：94℃/5 min→94℃/45 s,38℃/1 min,72℃/1.5 min, 45 cycle→72℃/10 min→4℃ forever, the effects of the compositions of RAPD-PCR reaction system on the gene difference of the cold and warm type of wheat were studied. An optimalzed RAPD system was set up： total volume was 25 μl,including DNA 20 ng template DNA,10×PCR buffer at 2.5 μl,dNTPs at 0.2 mmol/L,Taq DNA polymerase at 1 U,Mg^2＋ at 2.0 mmol/L,ddH2O12μl and 10 ng random primers.

关 键 词：冷型小麦 RAPD反应体系 稳定性 可靠性 

分 类 号：S512.1[农业科学—作物学]