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（摘要）

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。

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

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.