genomic library

tom tom_la at central.murdoch.edu.au
Fri Mar 6 06:43:53 EST 1998


Hi everyone

I'm currently screening a lambda gt11 genomic library, generated from a
partially digested bacterial genome, using a monoclonal antibody.  The
library is about 10^5pfu/ml, amplified to 10^8pfu/ml.  I have tried
several plaque-lifts but have been unsuccessful.  What I also noticed is
that plaques failed to form under the IPTG saturated filters, although the
plaques outside of the filter region (ie edge of the LB plate) grew well.
Does anyone who has used the gt11 system have any suggestions ?  Your help
will be greatly appreciated !

Also, if I incubate the LB plates containing IPTG (with the inoculated top
agarose) at 37oC until plaques form, place the filter on the plate, and
then further incubate for a couple of hours, will this be sufficient for
protein transfer onto the filter ?  

Thanks for your help in advance.

Tom  

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Tom La, PhD Student

Department of Microbiology
Division of Veterinary and Biomedical Sciences
Murdoch University
Murdoch, Western Australia, 6150

Tel :  61 08 9360 2247
       61 08 9360 2216
Facsimile :  61 08 93104144
Email :  tom_la at central.murdoch.edu.au

When ... everything goes against you ... never 
give up then, for that is just the place and 
time that tide will turn.
--- Harriet Beecher Stowe ---

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