cloning pGEMT - (May/12/2008 )

I'm working with pGEMT system of Promega for my cloning strategy and I have a question.
I want to know if it is possible transforming E.Coli with pGEMT for keeping this plasmid and then extract it and use it for cloning my PCR product or this is impossible and the only way to use the plasmid is buying the kit again.

Thanks a lot.

Ana.

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The pGem-T plasmids are linearized and then a T added using Klenow. If you manage to transform cells with it, it will be circularized plasmids only and will not be of any use to you. You can make your own T plasmid but transforming cells with the linearized form will not work. Sorry =)

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I supposed that, but I'm very principiant in cloning, so for that reason I want to be sure. Anyway thanks for your reply.

But now I have another answer, how can I make my own T plasmid, because I have some bacteria transformed with pGEMt circularized and I don't know how to do it.

Again thanks for your colaboration.

Ana

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I assume you meant another question, if so..

Check results Nos 1, 5 and 7.
http://search.vadlo.com/b/q?sn=158621799&a...ector&rel=0

..

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Yes, you are right I had another question...

Thanks for the protocol....

Ana

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