sequencing gel won't stick to Whatman paper

mullen at SMALL3.jsei.ucla.edu mullen at SMALL3.jsei.ucla.edu
Tue Jun 18 12:30:25 EST 1996


In article <4q68ad$g3v at highway.leidenuniv.nl>, <jasper at ruly46.medfac.leidenuniv.nl> writes:
> Path: nnrp.info.ucla.edu!info.ucla.edu!newsfeed.internetmci.com!in1.uu.net!EU.net!sun4nl!surfnet.nl!highway.leidenuniv.nl!jasper
> From: jasper at ruly46.medfac.leidenuniv.nl (Jasper Saris)
> Newsgroups: bionet.molbio.methds-reagnts
> Subject: Re: sequencing gel won't stick to Whatman paper
> Date: 18 Jun 1996 12:43:57 GMT
> Organization: Leiden University, The Netherlands
> Lines: 48
> Message-ID: <4q68ad$g3v at highway.leidenuniv.nl>
> References: <v01530500ade51e3a6d3d@[142.20.8.21]> <Dsy413.89I at ncifcrf.gov>
> NNTP-Posting-Host: ruly46.medfac.leidenuniv.nl
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> 
> Paul N Hengen (pnh at kaylor.ncifcrf.gov) wrote:
> : jlight at SICKKIDS.ON.CA wrote:
> 
> : : After I run a sequencing gel (using 35S-dATP for labelling) I have a lot of
> : : problems getting the gel to stick to Whatman paper.  I don't fix the gel
> : : after the run, because I've found that it's not necessary to get good
> : : autorads.  Unfortunately, and perhaps because I skip the fixing step, my
> : : gels don't always stick when I overlay them with Whatman paper 
(sometimes
> : : they do, sometimes they don't).  I've tried spraying the gel with water 
and
> : : blotting off the excess with Kim wipes prior overlaying with Whatman, 
which
> : : helps sometimes, but it's no guarantee.  Sometimes part of the gel 
sticks
> : : and part doesn't, so the gel ends up ripping.  Advice from anyone who
> : : routinely transfers their gels without fixing would be greatly 
appreciated.
> 
> Hoi,
> 
> For several years now we don't fix the sequencing gels. Both plates were
> repelled (Pharmacia) and we used Whatman 3MM. For several months (>6 ) we
> have changed to S&S type 2 paper and Pharmacia changed their Repel to
> Repel ES (Extra Strong?). Some three months ago I also have had
> difficulties with gels sticking to the glassplates instead to the S&S
> paper, but mainly with 10% PAA 1/50 + Glycerol SSCP gels.
> 
> The repel has certainly effect during pouring, but I don't know for sure
> if it one of the variables in the sticking problem. The front, larger
> plate is now treaten less often than the smaller, back plate. Saves again
> time, but beware. 
>  I treat the paper a bit different now. Longer lying on the plate was not
> the help I hoped for. An over dessicated surface could be, but I proceed
> immediatly after dismounting the two plates. Pressing the back of the
> paper with the back of a razor blade did help better than waiting for
> several minutes (1-10min). Not flipping the paper-gel-plate set with the
> second plate, but just lifting the paper over itself worked the best for
> several gel types. 
> 
> Good luck with thread of advice, it is a lot to choose from
> 
> 
> Jasper
> 
> J.J. Saris
> Dept. Human Genetics
> Leiden University
> Sylvius laboratory
> wassenaarseweg 72
> 2333 Al Leiden
> 
> 
> 





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