separation of ssDNA from dsDNA

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

j.migeon at 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

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