Inoue Competent Cells

Amna Saeed asaeed at NEU.EDU
Fri Mar 22 23:33:25 EST 1996


Hi:
			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 ??????????
																																														Amna Saeed
																																														Dept. of Biology
																																														Northeastern University



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