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plasmid rescue (plasmids with incompatible ORI)


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#1 geetika

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Posted 07 July 2009 - 08:18 PM

Hi,

I'm trying to ligate two plasmids (one with kanamycin and other with ampicillin selection markers) containing similar (pMB1 and ColE1) ORI's. I think as they fall in the same plasmid incompatibility group, I don't get any transformants on LB Kan/Amp selection plate. I'm using DH5 alpha chemically competent cells for transformation. Can someone please suggest if there is any bacterial strain where in a plasmid with two similar ORI's can exist? Or is there any other way to get this work done.

Thanks a lot

#2 jiajia1987

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Posted 07 July 2009 - 10:08 PM

I don't think there is a way to get two plasmids of the same ORI to exist together..

#3 molgen

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Posted 08 July 2009 - 07:31 AM

Instead of reinventing the weal, ask around if anyone has a pCR vecter from Invitrogen. It has both kanamycin and ampicillin resistances.




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