Separating ssDNA from dsDNA (asymetric PCR) ???
Mr. J. Sasse
jsasse at hgmp.mrc.ac.uk
Wed Oct 11 12:48:16 EST 1995
Two column based systems are available commercially to separate ssDNA from
dsDNA after PCR. The SSAM PCR purification system from Clontech and the
Centriflex PCR purification kit from Advanced Genetic Technologies Corp.
(agtc at access.digex.net) supplied in the UK by nbl Gene Sciences. The
latter seems to be a lot simpler to use and it is the only one I tried
and it works very well.
You just have to add the supplied PCR binding buffer to your finished PCR
reaction; transfer the aqueous portion of the sample to the Centriflex
cartridge; spin for 5 sec. in a microfuge which binds the protein and the
ssDNA to the membrane; add TE to elute the ssDNA and spin again for 5 sec.
Thats all, hope it helps
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