lane tracking

Eugen C. Buehler buehler+ at pitt.edu
Wed Apr 10 16:15:32 EST 1996


Ron,

   If your 377 gels have a consistant migration of the lanes then
something is definitely wrong.  The most likely cause of this would be a
heterogeneous gel.  To avoid this problem, I'ld make the following
recomendations

   1.  Use Long Ranger=81 gels.  They really do improve your results over m=
aking
         your own polyacrilamide mix.

   2.  Mix everything completely.  Too many people think that the gel will =
set
         if they don't pour seconds after adding the TEMED.  Mix a little l=
onger
         than you think is nescesary before pouring.

   3.  Look for patterns in the lane migration.  A consistant pattern might
         indicate an electrical problem (like a bum anode or cathode).

   Good luck, and make sure if you solve the problem to let everyone in
this group know how you did it.  These forums don't work if we don't share
answers as well as questions.

Eugen Buehler
buehler+ at pitt.edu

In article <4kfm5d$q5g at net.bio.net>, Ron Lundstrom <ron at eri.uchsc.edu> wrot=
e:

> Our lab is spending far too much time tracking lanes. We retrack more
> than half our gels. Hence they want me write lane tracking software,
> which I know won't be easy or short.
>=20
> ABI's lane tracking software does fine when the lanes run straight.
> In a previous job, I remember that some changes were made in the
> chemistry or gel preparation protocol that made things run straight.
> It wasn't 100%, but it helped a lot.
>=20
> Unfortunately, (`cause I'm the computer guy) I don't remember
> what was done. Can anybody give me some ideas on what to try?
>=20
> (We run ABI 377's with a 32 well comb.)
>=20
>      Thanks,
>      Ron L.
>=20
> --=20
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