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  

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

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：《安徽农业科学》2011年第17期10164-10166,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]探讨高简并性引物扩增庞大基因家族基因的特殊方法。[方法]采用高简并引物对鲤鱼基因组DNA分别进行了常规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]采用常规PCR仅得1条明显条带,得到的基因较少;采用常规降落PCR仅得到弥散性扩增,无明显条带出现;而采用小温度范围不规则跳跃降落PCR则得到了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的优化与选择提供了理论依据。[ Objective ] The aim was to explore the special methods for amplification of large-family genes by using primers with high degener- acy. [ 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 electrophoretie 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.

关 键 词：高简并性引物 不规则降落PCR 基因家族 扩增 

分 类 号：S132[农业科学—农业基础科学]