help needed on gel retardation assay

[Martin Leach]

In article <2mvop3INNcgp at dns1.NMSU.Edu>, twei at kirk (Tao Wei) wrote:

> Dear netters:
> 
> I am a graduate student and working with tomato plant.  I am running into  
> a problem in gel retardation experiments.  The DNA probe (200 bp prepared  
> by PCR) and crude nuclear proteins seemed to form very large complexes  
> (because of non-specific interaction?), stuck on the bottom of wells and  
> could not get into the gel.  The condition of binding reaction was pH 7.5,  
> 50 mM NaCl, 9-11% glycerol and some competitor DNA.  Does anybody have any  
> suggestions to help me out? Thank in advance.


How much protein??? Possibly too much!!

Try cutting down on the protein!

Martin
  

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.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
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