Protocol Online logo
Top : Forum Archives: : Molecular Biology

Gel separation of two fragments of similar size - (Sep/29/2001 )

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??



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