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HELP! How do you purify a 150bp PCR product for sequencing

baguet jbaguet at cri.ens-lyon.fr
Fri Dec 22 08:26:47 EST 1995


In article (Dans l'article) <DJy1It.D6s at uns.bris.ac.uk>, Barry Vipond
<vipond> wrote (écrivait) :

> Dear All,
> 
> I'm trying to purify a 150bp PCR product for automated sequencing. I've got
> a fair number of samples, so I don't want to gel purify. I've tried silica
> (Home-made), but the efficiency is very low (bigger products are fine). Glass
> fog or Mermaid, I assume will co-purify primers too.
> 
> What do I do? Does anyone have any nice recipes/methods that they've tried
> under similar circumstances? Perhaps there is a better binding buffer for
> the silica - I was using Guanidinium Isothiocyanate.
> 
> Hope someone can help.
> 
> Thanks
> 
> Barry (vipond at bsa.bris.ac.uk)

I use a protocole that permit direct sequencing on agarose gel
 slices without purifying DNA. All you need is low melting point
agarose. See Khorana et al, N.A.R. 22, 3425-3426



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