BIIIIG problem with top-agarose

CK Wen cwen at aux.btny.purdue.edu
Mon Nov 6 09:16:45 EST 1995


R:
Hallo reader

I am screening a genomic lambda libary. I am using host cells without 
selection.
The problem is that as I am transferring the plaques to Hybond N+ 
membranes,
the top-agarose adheres to the membrane thereby ruining everything.
I suspect this to happen because of moisture between the top-agarose and 
the medium.

So, what can I do to prevent this? I am cooling the medium to 50 deg. 
celsius before
preparing the plates so too hot medium should not be the problem.

Please  help me!

My e-mail address: hilde.nilsen at unigen.unit.no


------------------------------------------------------------------------
I guess you need to cool your phage plate at 4C for several hours before
lifting your phage to Hybond menbrane.  When the top agar is cold, it is
more solid, and it's the way I always do.

Good luck!

Chi-Kuang





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