Taq inactivation

John Dixon jpcd100 at mole.bio.cam.ac.uk
Wed Jul 26 05:36:23 EST 2000

In article <eXQZAUAiNYf5EA2i at demon.co.uk>, Dr. Duncan Clark
<drc at nospam.demon.co.uk> wrote:

> In article <397DC4E4.3259 at le.ac.uk>, A.F. Simpson <AFS7 at le.ac.uk> writes
> >Have you tried a simple phenol/cholorform extraction and ethanol
> >precipitation rather than using the purification kit?  Then you won't
> >have to worry about residual Taq or about buffer compatability.
> There has been another thread running on this and basically
> phenol/cholorform extraction and ethanol precipitation will not fully
> remove all the Taq.
> Duncan

Does anyone know the MWT of Taq off hand? We tried using S-300 spin
columns to purify PCR products after amplification and again after
digestion, hoping that the enzymes (Taq first and then the restriction
enzymes) would be retained in the sephadex, and the fragments would be
OK for ligation - we had mixed results. I asked amersham via their
website about this but got no response. Any clues?



John Dixon                    Lab 44 (1223) 334131
Wellcome/CRC Institute        Fax 44 (1223) 334089
Cambridge University
United Kingdom CB2 1QR       e-m: jpcd100 at mole.bio.cam.ac.uk

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