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Methods - 50 ssDNA fragment (Aug/09/2005 )

Good morning all!
Is it possible to ligate a ssDNA in a vector? Thanks in advance!


I don't think so, unless you make it dsDNA ...



Yes. To do this you need to cut your vector with 2 RE with one that gives 3' overhangs and one that give 5' overhangs. You then need to designed your ssDNA so that it has compatible ends to the 2 RE that you used. Ligate and transform as normal. I almost forgot- you will also need a 5' phosphate on the ssDNA.


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-Daniel Tillett-