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通过研究转化实验中加入质粒DNA的量、大肠埃希菌感受态细胞制备和质粒转化过程中冰置时间长短对感受态细胞最终转化效率的影响,分析其最佳转化效率参数,优化感受态制备和转化方法.结果表明,加入质粒(pBR322)DNA量为1～10ng,感受态制备过程中菌液冰浴时间在30～60min,以及感受态细胞加入质粒DNA并混匀后的冰置时间为30min和120 min时,其转化效率达到最高,在此范围之外,感受态的转化效率下降.本研究结果可为有高转化率要求的克隆转化实验提供有用的技术基础.
Abstract：The affecting factors on transformation efficiency of E.coli were studied,which include plasmid DNA dosage putting into transforming system,freezing time during preparation of competent cells and plasmid transformation.These tests were used to analyse the best transforming parameters and optimize the methods of preparation of competent cells and plasmid transformation.The results indicated that the parameters of the highest transformation efficiency in this study were 1～10 ng of plasmid(pBR322) DNA dosage,30～60 min of freezing time during preparation of competent cells,as well as 30 min and 120 min of freezing time during plasmid transformation.Out of these conditions,it decreased the transformation efficiency of E.coli.The results of this study will provide technology basis for the transformation tests requiring high transformation efficiency.
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基本信息:
DOI：
中图分类号:Q785
引用信息:
[1]罗锋,余腾,范丽梅.感受态细胞制备和质粒转化冰浴时间对大肠埃希菌转化效率的影响[J].江汉大学学报(自然科学版),2011,39(03):82-85.
基金信息:
武汉市科技局青年科技晨光计划项目(200950199019-03)