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Natural Science Forum / Biology / Biology / November 2003Why DNA can't move out of the wells in agarose electrophoresis
Hello,everyone, I am working on extraction of recombined Fosmid DNA,using alkaline lysis and ethanol precipitation.However,the isolated DNA remains in the wells and does not move even under the condition of 0.5% agarose gel, 200V,3hrs. Why and what can I do? Thanks a lot sherry > Hello,everyone, > I am working on extraction of recombined Fosmid DNA,using alkaline [quoted text clipped - 4 lines] > Thanks a lot > sherry dear sherry, the DNA loaded into agarose gel wells is mixed properly in what is called a loading bufer. this typically contains a tracking dye to track the DNA mobility and a high density compound such as glycerol. this glycerol increases the density of the dna to an extenct greater than the running buffer. so DNA is sunk into the wells. the same strategy is applied in Protein gels (PAGE) too. i think this info helps. with regards, santosh.. |
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