Weird problem in invitromutagenesis - (Apr/16/2011 )

Hi,

Iam doing invitro mutagenesis for a 5.2Kb plasmid using stratagene kit. I did not get any results with my plasmid, so checked the control in the kit. Unfortunately the control is not working. I have tried many times, but got no colonies. The conditions used were according to the instructions given by the manufacturer.
95°C 50 sec
95°C 50 seconds
55°C 1 min

68°C 5 minute x 18 times
68°C 5 minutes.

I even tried adding DMSO once. Nothing worked.

Any help would be greatly appreciated.

Regards,

jaya

Just a guess -- your problem might be in transformation. Have you checked your competent cells?

phage434 on Sat Apr 16 13:27:31 2011 said:

Thanks for the reply, but my competent cells are ok. Transformation with the other DNA of my worked well. Im not sure of the problem.

regards,

jaya

can you shortly comment on your control experiment?
you used the pWhitescript control plasmid ...or you talking about the pUC19 control?

Regards,
p