PAGE not uniform migration - (Feb/13/2008 )

Hello everybody,

I've a problem during PAGE of my cDNA-AFLP sample: the migration front is nor uniform, it make a curve. The outer samples seems to migrate slower than the inner....

I use TBE 1X (upper buffer) and TBE1X and NAoAC as migration buffer (down buffer)
The migration si made at 60watt during 2h
I don't make a pre run but I heat the upper buffer to 50°c before loading my samples.

I don't understand what happend...

Thks for your Help!!

there is always some curvature when one runs a gel. The gel smiles at you. Sometimes this can be due to changes in buffer in each sample.

by not prerunning you have not preheated the gel. the sides are cooler than the center and slows the migration.

you need to even out the heat distribution on the gel by prerunning and/or mounting the gel on a heating (cooling) plate.

1. Run the gel in the refrigerator
2.use a low voltage until all sample enter into the gel, then rise the Voltage

Running it slower gets rid of the smile too. Probably becasue warming is more uniform that way