Cloning in pGEM-T

Neil Saunders saunders at bio.vu.nl
Wed Nov 24 17:37:50 EST 1999


In my experience with pGEM-T, PCR product purification is not necessary if
you have a nice clean, specific PCR reaction (one major band on the gel).
If you clone direct from the reaction mix, you sometimes get one or two
contaminant inserts, but these occur at much lower frequency than the main
product and of course are easily distinguished on digestion

Neil Saunders

--
Department of Molecular Cell Physiology,
Faculty of Biology,
Vrije Universiteit,
De Boelelaan 1087,
1081HV, Amsterdam,
The Netherlands

tel: +31 20 4447194
fax: +31 20 4447229

email: saunders at bio.vu.nl
http://members.xoom.com/paracoccus


Christophe Jacob <jacob at scbiol.u-nancy.fr> wrote in message
news:jacob-2411991849160001 at betula.scbiol.uhp-nancy.fr...
> Does anyone know if the purification of PCR fragments (from agarose gel or
> with a column to eliminate other bands and/or oligonucleotides) before
> cloning in T/A vectors is a critical step ??
>
> Thanks for yours answers.
> Regards,
>
> Christophe.
>
> --
> Christophe JACOB
> Lab. Forest Biology
> University Nancy 1
> BP 239
> 54506 VANDOEUVRE
> email: jacob at scbiol.u-nancy.fr






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