How can I create a T vector for PCR product cloning - (Mar/19/2008 )

I have a pcr product apmplified with Platinum HF whihc leaves an overhanging A at the 3' end. Which enzyme should I use to cut my vector that leaves me a compatible end with my pcr product??> Thanks

T-overhang vectors are commercially available. My favorite is pGEM-T-Easy, available from promega. I actually made a T-overhand vector myself once but it takes time to do this and in your case you would probably be better off just purchasing it. It is available here: http://promega.com/catalog/catalogproducts...roductleaf_1039
They offer pGEM-T and pGEM-T-Easy which differ mainly in that pGEM-T-EASY has two EcoRI sites; one on each side of the MCS.

If you are trying to put your product into an expression vector you should probably amplify the sequence with primers containing RE sites appropriate to the expression vector.

Follow this thread to decide if it's worth purchasing or DIY.

http://www.protocol-online.org/forums/inde...?showtopic=6180

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