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
0
3 replies to this topic
#2
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Posted 16 April 2011 - 05:27 AM
Just a guess -- your problem might be in transformation. Have you checked your competent cells?
#3
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Posted 16 April 2011 - 06:02 AM
phage434, on 16 April 2011 - 05:27 AM, said:
Just a guess -- your problem might be in transformation. Have you checked your competent cells?
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
#4
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Posted 16 April 2011 - 07:48 AM
can you shortly comment on your control experiment?
you used the pWhitescript control plasmid ...or you talking about the pUC19 control?
Regards,
p
you used the pWhitescript control plasmid ...or you talking about the pUC19 control?
Regards,
p
