Biotinylation of dsDNA

Jeff Smith jmsst88+ at pitt.edu
Tue Jun 20 15:38:33 EST 1995


In article <3rmmi0$9n at hippo.shef.ac.uk>, M.C.Behrendt at shef.ac.uk
(M.C.Behrendt) wrote:

> Could anyone tell me how to biotinylate dsDNA (other than mixing the 
> dsDNA with photobiotin and puting it under a 400W UV lamp) 
> that is both easy and cheap.  I need to have approximately every 80th 
> base biotinylated.  The DNA is from any old plasmid that happens to be 
> lying about.

Unfortunately, I have only another question in response to your post.  I
guess that you have had some experience(and success?) with photobiotin. 
Besides looking for a red pellet following EtOH precipitation, are there
any other means of detecting successful biotinylation?  I am presently
attempting to dot blot my nucleic acid onto nitrocellulose and "probe"
with conjugated avidin-HRP for detection by ECL.  Any other suggestions?



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