vurquidiI at molbiol.ox.ac.uk
Mon Jan 15 10:56:04 EST 1996
In Article <30F63332.4635 at nott.ac.uk>, Simon Dawson
<Simon.Dawson at nott.ac.uk> wrote:
> I had an email from someone regarding the use of formaldehyde (formamide?)
>sequencing gels to get round this problem. Has anybody else heard of this
>method cuz I cant seem to get in contact with the person who suggested this
>method via the email address they gave me.
This method appears in Current Protocols in Mol. Biol.:
4 6 8
Urea (g) 42 42 42
38% acryl/2% bis (ml) 10 15 20
10X TBE (ml) 10 10 10
Deionized formamide (ml) 40 40 40
H20 (ml) 10 5 0
TOTAL VOLUME 100 100 100
(do not heat gel solution above 55oC)
Filter through Whatman No 1 paper
The gel sol. is viscous. Hold plates nearly vertical when pouring. Gel
should polimerize within 30 min.
Run the gel 45-70W constant power. This requires 60% higher voltage than
normal gels. (in my experience, more than 2500V, so be sure you have a good
power supply) DNA migrates about half as fast.
Fix gel in 5% acetic acid/20% methanol (v/v).
More information about the Methods