Hi
The pic attached is of a 1.3% agarose gel stained with gel red and run at 100V for 1 hour. The electrodes are approximately 30cm apart so this is rough 3 v/cm. I only loaded 3ul of ladder compared to the recommended 5ul as i thought this might be due to overloading the gel. I have had a similar problem with a different ladder so it isn't the ladder.
Any ideas?
thanks
james
bio.lonza.com/.../Lonza_BenchGuides_SourceBook_Section.
https://bio.lonza.com/.../Lonza_BenchGuides_SourceBook_Section.
Use fresh TAE buffer every time and maintain low temperature during running your gel because TAE buffer have low buffering capability and susceptible to high temperature and it is one of the important reasons of smiley bands.
you can get this effect when the ion concentration of the sample is very different from that of the buffer in the gel it is running in,,,for instance if the coloured dye is in 10x buffer and you use too much of it when adding to the size standard to get a good colour then the standard buffer concentration would be wrong
I think your gel is not even.In the corner side, it becomes thick and uneven that hampers ladder running.
It is about your gel. Please let it polymerase a little bit more. At least 30 min. Moreover if you are using a gel that is prepared one day before and stored do not use it.
The problem in gel preparation and the high voltage may cause W shape of the ladder, prepare fresh gel, and use a prechilled buffer or reduce voltage or use the ice pack.
Good luck
thanks guys i loaded 1ul of ladder instead of the 5ul recommended and i loaded the ladder after the samples and it seems to have solved the problem.
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