I am running a DNA PAGE after PCR (samples 6-15 are run in duplicate with the second sample digested) to determine serotonin genotypes. The ladder (well 1) is on the far right of the attached image). I would greatly appreciate any advice on how to enhance band brightness and definition, thanks.
Additional information: 5 uL ladder added, 10 uL PCR product per well, PH of the buffer is correct. Temperature of the room ~75F with Gel container NOT on ice.
These samples look like either too much salt or the gel is being run at too high a voltage . I would try running 7ul of pcr product and half the voltage that you are using. The lower voltage will also keep the gel cool minimising thermal diffusion of the sample and keep a thinner band of dna. Make sure that the gel is completely polymerised and be sure to wash out each well with running buffer before loading the samples
yes it would help or running the gel in a refrigerated room and even storing the running buffer in a fridge just before use but it is usually easy to lower either voltage or current on the power supply
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