3 prime fill in reaction?

Peter Ashby p.r.ashby at dundee.MAPS.ac.uk
Thu Sep 27 06:47:51 EST 2001

In article 
<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.


Peter Ashby
Wellcome Trust Biocentre
University of Dundee
Dundee, Scotland
Reverse the spam and remove to email me.

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