Costar Spinex vs Amicon micropure

[David L. Haviland, Ph.D.]

In <garrisons.6.000F870A at grace.cri.nz> garrisons at grace.cri.nz writes:

> I'm looking to purify smallish (118bp) PCR products from agarose gels for 
> sequencing. I am looking at Costar spinex tubes and Amicon micropures to do it 
> with. I haven't used either of these products, from the info I have I 
> understand that with the spinex the DNA goes right thru the filter, leaving 
> the agarose on top, This would leave a (possibly) dilute sample in the gel 
> buffer. The amicon micropure seems to concentrate the sample and allow for 
> changing the buffer.
> 
> Have i got it right? Has anyone used these products for purification of pCR 
> products for sequencing, and what are the results? All advice/hints appreciated


Brian:

Yes, we have used the Spin-X columns.  So far everything has been 
"ligate-able" and the largest restriction fragment that we've taken through 
has been 6 kb.  Others in the lab have tried PCR products though not as 
small as 118bp.  The volume of liquid is proportional to the starting 
volume of liquid inthe gel when applied to the column so after spinning one 
could do a a few dry butanol extractions.  We always (!) further purify the 
fragment with 2 phenol/ 2 phenol-clfm/ 2 clfm extractions followed by 
either NH4-Ac/EtOH pptns or if the fragment is big enough, a PEG pptn.  
As I've said, what we obtained has performed well in ligations and labeling 
as probe.

Hope this helps,
David

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