USS DNA & mutagen. THANKS!!
dnicker at vax.oxford.ac.uk
dnicker at vax.oxford.ac.uk
Sun Jun 6 16:36:13 EST 1993
A quick note of thanks to those who have replied to my posting
regarding USS DNA and mutagesesis (Patricia, Jim, Chris, Angelo and especially
Andre for the book :) ). All information was helpful, and thank-you to you
all for taking them time.
The protocols I received are too lengthy to include, but I learned the
following:
- important to keep multiplicity of infection MOI low when growing U
phage in order to ensure high uracil incorporation
- multiple (2) infection cycles will also greatly improve uracil
incorporation (ie- infecting CJ236 with Uphage)
- the background on neg. controls was due to self-priming (small phage
DNA fragments priming complementary strand synthesis). Although this is
undesirable, it should not be unexpected.
Thanks again!!
Darren Nickerson
Oxford
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