BIIIIG problem with top-agarose

zinc zinc at
Sun Nov 5 20:10:18 EST 1995


In article <47fj07$n4v at>,
Hilde Nilsen  <hilden at> wrote:
>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.

i have two suggestions for you.

1) try to use plates that have been 'cured' for a day or two so that
they are not to wet when you apply the top agar.

2) this is what i usually do to prevent the top agar from lifting off.
i just cool the plates to 4 deg C after the plaques have grown and i
have not had any problems with it lifting off when i do this.

- -pjf

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