High Efficiency Transformation (libraries)
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
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.
More information about the Methods