Agarose gel electrophoresis problems
Posted 24 April 2009 - 03:39 AM
Posted 24 April 2009 - 04:15 AM
Please help! I am having a problem running my agarose gels. The ladders always end up looking very distorted. The bands are smeared towrdas the top end, but fairly well defined at the bottom end. The middle looks like all the bands have landed on top of each other. I have tried different voltages from 80-120V and different buffers (TAE and TBE), but to know avail. In the attached image, I tried a series of different ladders to see if it was the particular ladder I was using. Does anyone know what the problem could be?
it looks to me like your ladder is degraded.. how old is it? also do you change tips every time you dip in (EtBr will degrade the DNA)? Do you aliquot it first (this protects against the effects of freeze-thaw)?
Posted 24 April 2009 - 04:49 AM
Are you sure you're casting the gels in the buffer you're running them in, and not in water? Are you sure the buffer is the right strength?
Do you have another power supply you can try?
Is the power supply set to constant voltage or constant current?
Do you have another gel electrophoresis chamber you can try?
Posted 24 April 2009 - 07:54 AM
Posted 24 April 2009 - 08:39 AM
It might be a density problem related to this or the agarose not being melted properly.
Posted 24 April 2009 - 10:35 AM
genius does what it must
i do what i get paid to do
Posted 24 April 2009 - 09:00 PM
Posted 25 April 2009 - 03:38 PM
Please stirr after every few min of boiling youer agarose and they will disappear.
Best of luck.
Edited by nirajm, 25 April 2009 - 03:38 PM.