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Gel separation of two fragments of similar size


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#1 anonymous

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Posted 29 September 2001 - 09:00 PM

I am trying to resolve two restriction fragments of nerly the same sizw. However, even after i ran the sample to the very end of the gel, the DNA fragments appeared as one band. What can I try to resolve it? And why do you think it will work??

Thanks.

#2 anonymous

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Posted 29 September 2001 - 09:00 PM

You can resolve the bands on a 8% acrylamide gel run in 1X TBE. Make this gel exactly as you would an SDS-PAGE gel, but leave out the stacking gel. Stain using conventional Ethidium bromide and visualize after including a mandatory 15 minute destaining in water. Alternatively, include 0.1% Low melting Seakem GTG in standard 1.0% regular agarose gels. For all fine separations, use TBE and not TAE as running buffers.

Good Luck

ChithraMycoBact Experts, IncA Division of DSTF-Global




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