In article <48pit6$2cl at infoserv.rug.ac.be>, nivil at gengenp.rug.ac.be says...
>>Dear Dr. Sonya Clark,
>I'm using this vector and i didn't get any results yet, not even in my
>controls following INVITROGEN instructions.
>I would appreciate if other people can comment on this.
I hope you are using the bacteria they supply with the vector (one of their
TOP10 derivatives, I think) as this has the necessary gyr+ phenotype.
The reason I ask is that one of their reps told me that I could still use
DH5alpha as for other vectors and was surprised to hear that it wouldn't work.
As was discussed in the group earlier, most of the normal host strains are
gyrA96 and will not work with this positive selection protocol.
If its not as simple as this then I hope you find your answer soon.
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)