Cloning in pGEM-T

Neil Saunders saunders at
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

Christophe Jacob <jacob at> wrote in message
news:jacob-2411991849160001 at
> 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
> email: jacob at

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