Ligation Help Requested
Zophonias Oddur Jonsson
zjons at rhi.hi.is
Sun Mar 28 20:45:29 EST 1993
In <27MAR93.19921652.0075 at UNBVM1.CSD.UNB.CA> E85J at UNB.CA (E85J000) writes:
>I'm in serious need of some help! I have a PCR product with
>engineered NdeI cut sites at both 5' & 3' termini. I'm trying,
>without any success, to cut and ligate into the NdeI site of
I recognize your problems.
I had a similar experience when PCR-cloning with an engineered NdeI
site. Then I came across an article where it was suggested treating the
PCR product with proteinase K (and then phenol-extracting carefully of
course) before digesting with restriction enzimes. Unfortunately I can't
find the article at the moment so I can't give you the reference, but
anyway this worked out fine for me and I got the desired ligation.
I hope this information is of some use !
Zophonias O. Jonsson: zjons at rhi.hi.is
Department of Molecular Genetics,
University of Iceland, Reykjavik, Iceland.
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