SSCP PAGE problems

Francisco G da Nobrega ib - bio 7588 fgdnobre at usp.br
Tue Oct 17 16:18:23 EST 1995


On 16 Oct 1995, Ayo Ola wrote:

> I'm trying to screen some DNA samples for mutations using the SSCP=20
> method.  After running my  5% PAGE + 10% glycerol gel for 15hrs at room=
=20
> temp my gels have refused to stick onto the Whatman paper and every=20
> attempt to dry the gel ends up losing the gel. My gel is made up as ffws=
=20
>      16ml 50% glycerol
>       8ml  50% Acrylamide Bis(49:1)
>       4ml 10X TBE
>       0.48ml 10% APS
>       0.08ml TEMED
>       51.44ml dH20
> Running buffer used is 0.5% TBE.
> I would be very grateful if anyone could help me out as I'm running=20
> crazy.
> Please mail replies, thank you.
>=20
> Ayo
>=20
Dear Ayo:

You could forget about drying and just remove one of the glass plates and=
=20
cover the gel with Saran. Put it inside on of the Kodak cardboard=20
cassettes (or the Sigma flexible plastic ones) over your Xray film with a=
=20
intensifying screen underneath and expose OVN. That's how we usually do=20
with the Maxam and Gilbert sequencing.=20

F.G. Nobrega
Universidade de S=E3o Paulo
Biologia - IBUSP
C.P.  11461
05422-970   S=E3o Paulo  -  SP  - Brasil
Voice: (011) 818-7588   Fax: (011) 818-7553    E-mail:  fgdnobre at usp.br
=09=09=09



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