DOP-PCR阴性对照中的产物分析及引物浓度的优化  

Product of Negative Control and the Optimization of Primer Concentration for DOP-PCR

作　　者：张伟伟[1] 张衍梅[1] 赵刚[1] 陈戟[1] 刘江东[1] 余其兴[1] 

机构地区：[1]武汉大学生命科学学院,湖北武汉430072

出　　处：《武汉大学学报（理学版）》2006年第2期220-224,共5页Journal of Wuhan University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30370772;30400044)

摘　　要：由于在DOP-PCR实验过程中,通常的防污染措施和对反应体系及条件的优化都无法完全消除阴性对照中出现的扩增产物,因此本研究对上述扩增产物进行了纯化、克隆和测序,发现该产物并非污染所致,而是由于引物自身的兼并性,序列相互配对而产生的引物多聚体;并且还对与该扩增产物产量密切相关的引物浓度进行了优化探讨,实验结果表明2.2μmol/L是减少该产物产量的比较理想的引物浓度.In DOP-PCR experiment, amplification product often generate in negative control. The measures preventing contamination in common and the optimization for reaction＇s system and condition can not erase this phenomenon completely. By sublimation,cloning and sequencing for amplification product, it is found that they are generated not because of the contamination but the degeneracy of primer. Thedegenerate sequences pair each other and generate the primer polymer. The concentration of primer is very important to the amplification product＇s yield. So the optimizing study on primer＇s concentration of reaction condition is carried on in this experiment as well. And the result shows that 2. 2 μmol/L is better eoneen tration to reduee the yield of amplifieation product.

关 键 词：DOP-PCR 引物多聚体 优化 

分 类 号：Q343[生物学—遗传学]