Self-made TOPO cloning

Marcus Wernitz viacx at mlucom6.urz.uni-halle.de
Fri Aug 28 03:21:50 EST 1998


Has anybody thought of a self-made TOPO-like-cloning Kit?

It should not be a problem to sythesize and ligate the
EcoRI-EcoRI-Fragment with the AflI-site and the
topoisomerase-binding-sites into any plasmid at hand. 

But does the buyable calf-thymus topoisomerase bind to the binding sites
like it was originally described for the Vaccinia topoisomerase?

Or migth there be a mojor problem in transformation of the competent
cells. like DH10beta?

Who is having any good ideas about this?

Yours,

Marcus




-- 
Diplombiologe
Marcus Wernitz
Martin-Luther-Universitaet Halle-Wittenberg
Landwirtschaftliche Fakultaet
Institut fuer Pflanzenzuechtung und Pflanzenschutz
Lehrstuhl fuer Phytopathologie und Pflanzenschutz
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Germany

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