High Efficiency Transformation (libraries)

the End jgraham at bronze.ucs.indiana.edu
Thu Feb 17 14:38:51 EST 1994


Thanks to Mike Coady and Rafa Maldonado for pointing out the Inoue 
transformation study. 

In growing cells at 18C prior to Ca treatment, has anyone arrived at a good
inoculation and growth period ? I hear a couple "overnight" references,
but I would prefer not to determine the doubling time at this temperature.
I'm going at 20-22C, room temp here.

I also came upon two interesting options, one to treat thawed competent 
cells for 10 min with 0.5 M B-mercaptoethanol (5 ul/ 200 ul cells) and the 
other involves a "deoR" mutation in a strain that might be more readily
transformable, presumably a DH5 derivative called INValphaF'. Any comments
on this mutation most appreciated.

Thanks again,

Jim
J. Graham




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