running ssDNA (30-1000nt) in denatured conditions
cutest_chemist from yahoo.com
(by cutest_chemist from yahoo.com)
Wed Nov 19 03:10:16 EST 2008
On Nov 5, 12:51 am, TC <tracyscie... from gmail.com> wrote:
> Hi all,
> I am trying to run DNA under denatured conditions. Has anyone run single
> stranded DNA (30nt to 1000nt) under denatured conditions.
> I am trying to decide whether to run a low percentage urea acrylamide gel or
> an alkaline agarose gel. Any thoughts/ideas? I am thinking maybe running a
> low percentage (3-4%) urea acrylamide gel, but wonder that it may not be
> able to resolve ssDNA above couple hundred nucleotides in length even in low
> acrylamide such as 3-4%? Any thoughts? Has anyone run alkaline agaorse for
> ssDNA 30nt-1000nt? If so, may I ask for your conditions?
I m currently using 20% Polyacrylamide gel(PAGE)for ssDNA (20-30 nt)
and 15% PAGE for ssDNA of 30-50mer.Its giving good results...Hope this
will help u..For more lengthy strans i think u should try 8-10% PAGE.
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