crackling sequencing gels

T. Max M. Soegaard tmms at mbio.aau.dk
Fri May 26 13:21:29 EST 2000


Thanks alot for all this.
And I think I found the lever to pull to make my ng program stop posting
in unreadable formats. That is I hope so..
Is this o.k.?
-Max

"A.F. Simpson" wrote:
> 
> T. Max M. Soegaard wrote:
> >
> > thin 0.4mm sequencing
> > gels in a size of app 50cmx30 cm. Do you think it would hurt to
> > transfer them to filter paper first and then fix and wash them.? Im
> > thinking that it will be some very delicate handling steps with a big,
> > thin, gel..(but I dont know)
> 
> I didn't realise they weren't being fixed!  That'll teach me to make
> assumptions.  Yes, they _definately_ need to be fixed.
> 
> As for handling the gel:
> 
> The filter papaer would fall to pieces in the fix, and if it didn't,
> you'd have a piece of filter paper soaked in fix to dry out as well as
> the gel.
> 
> The way I used to handle fixing sequencing gels was to soak for 1 hour
> in 10% acetic adic/10% methanol (as per the Sequenase 2 kit booklet).
> However, this was for 1.2mm-0.4mm gels, so a shorter time for a 0.4mm
> gel should be sufficient.  I find pouring the fix into the tray and then
> lowering the gel in SLOWLY is much better than putting the gel in the
> tank then pouring in the fix.
> 
> To transfer to the paper I lift the gel out of the tank rather than
> pouring off the fix.  Again, SLOWLY is the key; if the gel starts to
> slide it can usually be rescued by tilting the plate back down.  Then
> transfer to a double thickness of filter paper.  The sheet in contact
> with the gel gets quit wet and therefore fragile, so a double thicknes
> makes it easier to handle.
> 
> (BTW, you _are_ posting in HTML.  Luckily my browser can read it and
> quote back properly, but some people's won't and they probably won't
> bother reading your posts.  I don't know anything about Mozilla, but
> there muct be an option somewhere to turn the HTML off.)
> 
> > -Max






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