Help! - EMS mutagenesis
brad at corona.med.utah.edu
Mon Jul 10 13:08:49 EST 1995
In article <T.Barnett-0907951236160001 at 18.104.22.168>,
T.Barnett at path.utas.edu.au (Tim Barnett) wrote:
> I am trying to mutate an endogenous alkaline phosphatase gene from
>Aeromonas sobria and intend to perform subsequent TnphoA mutagenesis on
I take it that you haven't found your mutant of choice yet. First a couple
questions and then a suggestion.
1) Are you sure that Aeromonas sobria has only one phosphatase? If there
are multiple phophatases that are active on LB/BCIP plates then the chances
of seeing a white is much lower.
2) Have you done a killing curve with your mutagenesis yet? This is a
handy thing to do in order get a handle on how hard you are hitting your
A realy good resource for your experiment is Jeffrey H. Miller's "A Short
Course in Bacterial Genetics" 1992. Cold Spring Harbor Laboratory Press.
ISBN 0-87969-349-5. Specifically, Experiment 3, Mutagenesis with EMS.
Brad Nicholson |"If it worked the first time, it wouldn't be
Department of Pathology | research."...Brad Nicholson
University of Utah | Live from behind the Zion Curtian.
Salt Lake City, UT 84132 |
brad at corona.med.utah.edu |
or: (801)-581-4365 | My opinions are solely my own.
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