Gels keep cracking
pxpst2 at unixs.cis.pitt.edu
Sat Nov 28 12:54:12 EST 1998
I routinely dry my 14% gels with about a 10% cracking frequency. My
samples use I125 as the label source. I do not fix the gels and I do not
use glycerol. But if you can get rid of the fixing step and dry the gel
immediately following the run you may have more success. The required
amplification may not be needed if you look for a "specialty film that is
more tuned toward S35.
I dry my gels with a uniform heting of the gels for 2 hrs @ 80 C under a
vacuum. Be careful not to strech the gel when laying it down on the filter
paper. I usually lay the gel on the filter paper from the bottom to the
top of the gel because the remenant stacker layer is really sticky and can
cause the gel to elongate at the corners.
Hope this helps
In article <email@example.com>, S.Mckay at sct.gu.edu.au
(S. McKay) wrote:
> I've been having some problems drying mini-gels. I'm using 14%
> SDS-PAG to separate my protein (which is 35S-labelled). Following this I
> have been fixing the gels in 7% Acetic Acid for 20', rinsing in dH2O and
> then an amplification step using 0.5M Salicylic Acid for 20'. The gels are
> then dried onto Whatmans 3mm Chromatography Paper under vaccum and heat on
> a BioRad Gel Slab Dryer for 1.5hrs. Less time than this and the centre of
> the gels do not dry. Unfortunately, I've been experiencing severe cracking
> problems with the gels which is obviously unacceptable.
> I am aware that the salicylic acid will remove much of the moisture
> from the gel but 1.5hrs seems a long time.
> I've also tried using the 0.5M Salicylic Acid combined with 2%
> glycerol but this makes no difference and if anything the cracking is
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