separation of ssDNA from dsDNA

Richard V. Giles giles at liv.ac.uk
Sat Sep 19 07:00:26 EST 1998


j.migeon at cerep.com wrote:

> If anyone knows how to effectively separate single-stranded
>
> DNA from double stranded DNA (no gels),  I would be grateful
>
> for any suggestions.
>
> Thanks,
>
> Jacques
>
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The classic method for this is chromatography on hydroxyapatite collumns.

Basically you apply your sample in a very weak phosphate buffer and elute
with increasing concentrations of phosphate buffer. Single stranded stuff
comes off at lower phosphate than ds DNA.

Detailed methodology in Maniatis "Molecular Cloning" ppE30-E34.

Also works well as a batch method if you spend the time setting up the
optimal concentration of phosphate so that only ds DNA binds the resin.

--
Richard V. Giles Ph.D.
School of Biological Sciences
University of Liverpool
Tel +44/151 794 4389
Fax +44/151 794 4349
http://www.liv.ac.uk/~giles/





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