Inoue Competent Cells
asaeed at NEU.EDU
Fri Mar 22 23:33:25 EST 1996
I also recently tried out the Inoue protocol, but since I didn't
have access to a cooling incubator, I grew up my 250ml culture
inocculated with about 10 colonies of SURE cells (Stratagene) in an
incubator at R.T. (thermometer read about 23oC). I inocculated at
10:30 pm and at 10:30 the next am, the cells were precisely at 0.6
O.D.600 ( I was surprised). I tried to calculate the transformation
efficiency of these cells (10ng plasmid DNA added after 10 min w/ 17mM
DTT) and got between 3.0 and 3.75x10E8 which I was quite happy with.
Perhaps 23oC rather than 18oC is not such a bad trade off even if you
lose some efficiency but reduce doubling time ??????????
Dept. of Biology
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