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Weird problem in invitromutagenesis


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3 replies to this topic

#1 jaya2020

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Posted 16 April 2011 - 03:34 AM

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.
95C 50 sec
95C 50 seconds
55C 1 min

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

I even tried adding DMSO once. Nothing worked.

Any help would be greatly appreciated.

Regards,

jaya

#2 phage434

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Excellent

Posted 16 April 2011 - 05:27 AM

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

#3 jaya2020

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Posted 16 April 2011 - 06:02 AM

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 pDNA

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Good

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




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