Slanted Bands on Agarose Gels

Will Fischer wfischer at bio.indiana.edu
Tue May 24 12:01:21 EST 1994


Christopher W Botka (cwbotka at MERCURY.SFSU.EDU) wrote:
: Has anyone experienced bands(DNA) that run slanted on an agarose gel? -
: i.e. the bottom of the EtBr stained frag. of DNA seems to be migrating
: faster than the top.  This phenomenon occurs with all samples on the gel
: including the ladder and conc. stds.  However, it only occurs occasionally,
: and we have not been able to figure out why, although not from lack of
: effort.  We have varied current,  gel set up time, buffer, agarose brand,
: electrical socket (strictly voodoo), to no avail...

: Any input would be appreciated.  The data is still there, but it is ugly!

: -CWB

There was a thread on this a while back (w.r.t. keeping bands sharp on
blots).  Seems the culprit is different buffer conc. between gel and
running buffer, generally due to volume loss while dissolving the agarose.

Try making the gel up to volume by weighing the gel components _before_
heating, then adding dH2O _after_ heating to bring the weight up.

It seemed to help _my_ gels.

--
Will Fischer				wfischer at indiana.edu

Department of Biology			Voice:	812-855-2549
Indiana University			Voice2:	812-333-2433
Bloomington, Indiana 47405 USA		FAX:	812-333-7922



More information about the Methods mailing list