Separate DNA fragments of same length

Mark Schweder mschweder at gnv.ifas.ufl.edu
Sun Feb 7 21:19:05 EST 1993


In article <1ksj7nINN5r6 at access.usask.ca>, lur at jester.usask.ca (Rui Lu) writes:
> Hi,
> 
>       Is there anyone knowing how to separate DNA fragments of exactly same
> length (about 150 bp) with different sequence? These fragments were isolated
> by PCR using degeneracy primers. I don't want to screen hundreds of colonies
> by DNA sequencing. Also, I don't want to lose many amplfication products.
> 
>       Any smart ideas?
> -- 
> Ray Lu
> lur at sask.usask.ca
> Ph:(306)966-4440/4442

Try Denaturing Gradient Gel Electrophoresis (DGGE). This protocol involves 
a polyacrylamide gel with an increasing concentration of denaturant 
(formamide and urea) from top to bottom. I have used it with RAPD fragments 
and it seems to increase the bands compared to an agarose gel by about two 
times. I don't have the specific protocol available to me at the moment but 
if you are interested write me back and I can point you to them.
-- 
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   Mark Schweder                | There are three things that smell like fish!
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