3 prime fill in reaction?
p.r.ashby at dundee.MAPS.ac.uk
Thu Sep 27 06:47:51 EST 2001
<Pine.SOL.4.33.0109261515090.22413-100000 at mcmail.cis.mcmaster.ca>,
Kyle Legate <legatek at mcmail.cis.mcmaster.ca> wrote:
> Michael's correct. Since DNA polymerization proceeds 5'->3' there is no
> way to fill in the BsrD1 restriction product, as this would necessitate
> 3'->5' polymerization. T4 polymerase (and unmodified Klenow fragment)
> possess 3'->5' exonuclease activity, which is why you see your end being
> chewed away.
yep, that's I thought thanks for confirming it. We'll either revert to
pcr or custom linker and recut.
Wellcome Trust Biocentre
University of Dundee
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