- 期刊
A Fast Method for E. Coli Transformation
並列摘要
We develop a fast, simple and efficient method for E. coli transformation. In this method, plasmid DNA are mixed with 100 μl ice cold competent cells incubated in ice for 1 min, immediately freeze in liquid nitrogen for 1 min, following with 37℃ heat treatment for 1 min. Then directly spread onto the selective agar plate pre-warmed to 37ﾟC and culture for 16-18 hr. In general, the transformation efficiency is up to 21.49×10^6 colony forming units/μg DNA, though the efficiency for large (8.1Kb) plasmid is lower. The advantage of this transformation method is that all the transformation processes can be achieved within 3 min. It is significantly faster than most of the reported methods that usually require 75-90 min. This method also has the potentiality to be applied to the high throughput screening by using 96-well-culture plates for transformation.
延伸閱讀
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