Weird problem in invitromutagenesis
Posted 16 April 2011 - 03:34 AM
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.
Posted 16 April 2011 - 05:27 AM
Posted 16 April 2011 - 06:02 AM
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.
Just a guess -- your problem might be in transformation. Have you checked your competent cells?
Posted 16 April 2011 - 07:48 AM
you used the pWhitescript control plasmid ...or you talking about the pUC19 control?