Q: Does blunting w/ Klenow make PNK phosphorylation obsolete?

David Micklem drm21 at mole.bio.cam.ac.uk
Sun Jan 5 11:44:30 EST 1997


In article <5aofsf$k5h at mserv1.dl.ac.uk>, Wolfgang Schechinger
<wgschech at med.uni-tuebingen.de> wrote:

>Does it? or does it not?
>
>Anny suggestions are welcome!
>
>Wolfgang

It doesn't.  PNK puts on a _5'_ phosphate.  Klenow will not affect the 5'
end of your DNA at all:  5' overhangs are _filled in_ by extending the
3'-OH of the other strand until the two strands are the same length.  3'
overhangs are chewed back from the 3'-end.

David

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