甘油
大肠杆菌
电穿孔
甘露醇
离心
转化(遗传学)
化学
色谱法
细菌
生物
生物化学
基因
遗传学
标识
DOI:10.1016/j.ab.2011.02.036
摘要
Traditional protocols for preparing Escherichia coli for electroporation are laborious and often deliver highly variable transformation efficiencies. Many laboratories resort to purchasing expensive commercially prepared cells. This article describes a simple method for producing electrocompetent E. coli by centrifuging bacteria through a glycerol/mannitol density cushion. The method is rapid and replaces tedious multistep procedures with two 15-min centrifugations. Standard cloning strains consistently produce more than 8×10(9)transformants/μg pUC18, whereas the strains TG1 and LE392 display efficiencies of more than 3×10(10)/μg DNA.
科研通智能强力驱动
Strongly Powered by AbleSci AI