随机ssDNA文库不对称PCR扩增条件的优化及回收效率比较  

Optimization of Asymmetric PCR Reaction System and Purification of Random Single-srtand DNA Library

作　　者：周晏丞[1] 曹立亭[1] 陈露[1] 马跃[1] 

机构地区：[1]西南大学荣昌校区动物医学系,重庆荣昌402460

出　　处：《四川畜牧兽医》2015年第11期33-35,37,共4页Sichuan Animal & Veterinary Sciences

摘　　要：本研究开展了SELEX技术中ssDNA文库不对称PCR扩增及回收条件的优化试验,结果得出:20μL不对称PCR反应体系中上下游引物的最佳比例为60∶1,最优循环数为30个循环,回收效率最高的方法为乙醇沉淀法。此条件下,随机ssDNA文库不对称PCR扩增出来的条带清晰、稳定、特异性高,并具有较好的纯化效率。The asymmetric PCR amplification and purification methods of ssDNA library used in SELEX technology were optimized in this study. The result showed that the optimal number of cycles was 30,and the optimal ratio between forward and reverse primer was 60：1. Under above PCR conditions, clear and stable bands of ssDNA library were amplified. Ethanol precipitation method had the highest purification efficiency compared with the other two methods. This experiment laid technical foundations for selection of specific aptamers of target material in SELEX.

关 键 词：配基系统进化技术 ssDNA文库 不对称PCR DNA纯化 

分 类 号：Q78[生物学—分子生物学]