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!
Plant Science Laboratory | One of them is fish...
Agronomy Department | The other two...
University of Florida | Are growing on you...
Gainesville, Florida 32611 | (Frank Zappa, Jumbo Go Away)
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904-392-1890 | mschweder at gnv.ifas.ufl.edu
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DISCLAIMER: I know nothing about nothing.
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