(fwd) TA cloning : orientation problem

Shahram Mori smori at nmsu.edu
Tue Apr 18 01:32:45 EST 1995


> 
> Hi, I use the Invitrogen TA cloning kit and find that all the
> products land up in one orientation for one of my reactions.This
> causes headaches when I try and put it in an expression vector
> (the only one for this work ! )
>  Has anyone faced a similar problem ? any solutions ?
> Thanks
> NDF
I may be wrong but,
If all the products end up in one direction, there is strong indication that
the expressed PCR product is lethal to your competent cells.
I would try cloning the PCR in an expression vector with highly regulated 
promoters where you can control the expression of the gene.
Cheers
Shahram Mori					   _/\_
Program in Molecular Biology			  _\  /_
Dept. of chemistry and Biochemistry Box 3C	  \_  _/
NMSU  Las Cruces NM				    ||
88003






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