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Help!! w/Particle Purification and Resolution

Ed Rybicki ED at MOLBIOL.UCT.AC.ZA
Thu Dec 14 08:23:07 EST 1995


> How MIGHT i resolve different size things in this size range?
> 
> Any and all suggestions would be appreciated, no matter how crazy they may
> seem.
2 good methods...both OLD, but goodies:

1) agarose gel electrophoresis: if they are different, then they may 
have different charges, and will migrate differently in an electrical 
field.  And you can use just about any reasonable buffer (down to pH 
5 and up to pH 9), and down to 0.5% agarose.  Then simply press, 
airdry, and stain with Coomassie.

2) sucrose gradient zone electrophoresis: only thing we could use for 
preparative separation of two different aphid picornaviruses some 
years back; described in that wonderful bible of techniques for 
classical plant virology, Kado and Agrawal.

Both presuppose your particles are in fact from different viruses and 
not built from the same coat protein - in which case Percoll 
gradients or ENDLESS cycles of sucrose gradient centrifugation 
coupled with taking only the outsides of the peaks may work.

Good luck....

                     Ed Rybicki, PhD  
      Dept Microbiology     |  ed at molbiol.uct.ac.za   
   University of Cape Town  | phone: x27-21-650-3265
   Private Bag, Rondebosch  |  fax: x27-21-650 4023
      7700, South Africa    |   
    WWW URL: http://www.uct.ac.za/microbiology/ed.html      
                                        
"And then one day you find, ten years have got behind you"



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