transforming with RE digest

Michael G. Kurilla mgk2r at Virginia.EDU
Wed Jul 28 14:19:21 EST 1993


You do not need to heat inactivate in order to transform;
however, heat inactivation generally improves the outcome.
This is probably due to the enzyme remaining stuck to the DNA
(which T4 DNA ligase will do) and reducing the transformation
efficiency.  If you are going to transform by electroporation,
then you need to remove the salt from the reaction (or else
dilute way up) since any salt raises the conductivity of the
medium too high for electroporation.  Even after just a
ligation, we dilute 5 fold before transformation by
electroporation.  Heat inactivation also improves
transformation by electroporation (there are several NAR
articles that have documented this within the past several
years).

Mike Kurilla
University of Virginia
mgk2r at pcmail.virginia.edu



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