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Gel Electrophoresis Bowed Migration Pattern Troubleshooting - (Nov/25/2006 )

Howdy,

Trying to troubleshoot a DNA Fingerprint 1.5% agarose gel electrophoresis gel- got strange bowing migration pattern where the ends migrated faster than the center lanes. Same molecular marker was used in lanes 1 and 3 in the attached image, which leads me to believe it's a gel problem.

Does anyone have an idea of what might cause this? I've heard this not an entirely uncommon problem?


Thanks for any suggestions!!!

-vlk1968-

Some things to check:
* has the gel completely set? Perhaps the outside has set and the middle has not.
* are you running the gel hot? High current could cause heating most visible in the center.
* is the gel thickness uniform? A bowed gel tray could cause this.
* Are the electrodes straight and uniform and covered with buffer? Electrode contact at only the edges could cause this.
* Is the buffer covering the gel? Nonuniform buffer thickness could cause this.

-phage434-

In addition to what phage434 has pointed out, try to fill the empty wells with roughly same amount of loading buffer; be sure to reduce the ionic strength of your samples by gel filtration or dialysis method.

-genehunter-1-

Excellent ideas! Thank you guys for your quick and helpful replies!

-vlk1968-