U-shape bands in agrose electrophoresis

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U-shape bands in agrose electrophoresis

Postby JanChang » Jun 28 2016 9:06 am

1.5% agarose gel; dimension is 107mm(W)x60mm(L); running buffer 0.5x TBE; voltage is 100V, and the staining dye Gelred was added in gel directly. The bands are U-shaped as shown in the attached gel picture. This pattern had been observed a lot in my lab and not sure what happened. Any suggestion? Thank you.
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Re: U-shape bands in agrose electrophoresis

Postby relaxin » Jun 28 2016 3:34 pm

Your voltage is too high. Try run at 70 volts.
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Re: U-shape bands in agrose electrophoresis

Postby mchlbrmn » Jun 29 2016 3:50 am

It looks like it's a combination of that and the amount loaded. The bands with less DNA are straighter.
I wonder if 1xTBE would perform better under these conditions?
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Re: U-shape bands in agrose electrophoresis

Postby mdfenko » Jun 29 2016 12:34 pm

as mchlbrmn wrote, you have overloaded the gel.

also, you may be taking too much time to load or letting the gel sit before starting or both. this allows sample to diffuse into the gel surrounding the well and retarding peripheral migration.
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Re: U-shape bands in agrose electrophoresis

Postby 29yrsExperience » Sep 07 2017 6:45 pm

The substance you use as a sinking agent can also affect band shape. Ficoll tends to allow less distortion than sucrose, for instance. For the prettiest gels I usually stain after I run it.
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