sequencing gels; problems with plates and combs

daj (David Johnston) daj at nhm.ic.ac.uk
Fri Jan 21 03:46:32 EST 1994


On Thu, 20 Jan 94 21:29:48 GMT,
  Anthony J. Persechini writes:

2>For some time I have used a method for pouring BRL type wide sequencing 
gels in>which the short
>plate is treated with silane and the back plate is treated with a modified
>silane that causes the gel
>to be cross-linked to this plate. I then dry the gel right on the glass.
>Unfortunately, the gel sometimes (right now, always) sticks to the short plate
>also. Has anyone who uses this method determined the root cause of this problem?
>If so, what are the remedies?

How often do you re repel silane your short plate? A build up appears 
to cause sticking in our hands. Manniatis has protocols for stripping off 
old repel silane.

>Also, how can one avoid the problem of crumpling teeth on a sharks tooth comb?
>i.e; is there a nice relibale method that ensures that the comb will go back in
>easily?
>
Biorad used to sell metal sharkstooth combs, these may have been tougher 
(though at a price). I am not sure about the anatomy of the BRL system 
but on our IBI and Hybaid rigs we tend to inset the comb before putting the 
gel assembly onto the rig, ie before the plates/tank assembly is clamped 
together. This makes the comb easier to inset although you obviously 
have to be careful to not prise the plates apart - if you clamp plates 
with bulldog clips for casting, you can leave the lower ones in place 
whilst doing this for extra security.
Cheers,
DAJ

David A. Johnston
Dept of Zoology, The Natural History Museum, Cromwell Road,
South Kensington, London SW7 5DB.
(tel 071 9389297, fax 071 9388754, email daj at nhm.ic.ac.uk)



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