(fwd) TA cloning : orientation problem
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 ?
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.
Shahram Mori _/\_
Program in Molecular Biology _\ /_
Dept. of chemistry and Biochemistry Box 3C \_ _/
NMSU Las Cruces NM ||
More information about the Methods