CAJ | 학술논문

[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.
[목적]탐토료고간병성인물확증방대기인가족기인적특수방법.[방법]채용상규고간병인물대리어기인조DNA분석진행료상규PCR확증、상규강락PCR확증이급우화후적소온도범위불규칙도약강락PCR확증.[결과]채용PCR부득1조명현조대,득도적기인교소;채용상규강락PCR법부득도미산성확증,무명현조대출현;이채용소온도범위적불규칙도약강락PCR칙득도료득도료3조명현조대화다개기인,확증결과이상.[결론]위방대기인가족확증조건중적우화여선택제공료이론의거.
[Objective] The aim was to explore the special methods for amplificati,on 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.