Application of Abnormal Touchdown PCR with High Degeneracy Primer in Amplification of Large-Family Genes  

高简并引物小温度范围不规则降落PCR在庞大基因家族扩增中的应用(英文)

作　　者：华慧颖[1] 王芳[1] 常重杰[2] 杜启艳[2] 

机构地区：[1]中州大学化工食品学院,河南郑州450044 [2]河南师范大学生命科学学院,河南新乡453007

出　　处：《Agricultural Science & Technology》2011年第2期188-190,共3页农业科学与技术（英文版）

摘　　要：[Objective] The aim was to explore the special methods for amplification of large-family genes by using primers with high degeneracy.[Method] By using the primers with high degeneracy,conventional PCR,conventional touchdown PCR and the optimized abnormal touchdown PCR were respectively carried out to amplify the genomic DNA of Cyprinus carpio.[Result] Only one evident electrophoretic band and a few Sox genes were obtained by using normal PCR;no obvious electrophoretic band but dispersive product was obtained by normal touchdown PCR;ideal result was obtained by the abnormal touchdown PCR that three evident electrophoretic bands and much more Sox genes were amplified.[Conclusion] The research provided theoretical basis for the optimization and selection of PCR amplification conditions of the large-family genes.[目的]探讨了高简并性引物扩增庞大基因家族基因的特殊方法。[方法]采用常规高简并引物对鲤鱼基因组DNA分析进行了常规PCR扩增、常规降落PCR扩增以及优化后的小温度范围不规则跳跃降落PCR扩增。[结果]采用PCR仅得1条明显条带,得到的基因较少;采用常规降落PCR法仅得到弥散性扩增,无明显条带出现;而采用小温度范围的不规则跳跃降落PCR则得到了得到了3条明显条带和多个基因,扩增结果理想。[结论]为庞大基因家族扩增条件中的优化与选择提供了理论依据。

关 键 词：Primers with high degeneracy Abnormal touchdown PCR Large-family genes AMPLIFICATION 

分 类 号：Q78[生物学—分子生物学]