Ligase fidelity

Brett Lindenbach brett at BORCIM.WUSTL.EDU
Thu Dec 3 14:40:39 EST 1998


>I wonder if anyone has any information on the requirement of 100%
>complementary ends for the various ligases available? I would like to
>ligate a pool of ds DNA with random 4 base overhangs to an adaptor with
>specified 4 base overhang. Which ligase is the best for maximising
>ligation of 100% complementary overhangs only?
>
>Raj
>

The fidelity of T4 DNA ligase is actually pretty good, although some
numbers to back that up would best be found in the literature.  There
were many great papers written on this enzyme back in the early 70's,
so try looking up work from HG Khorana or CC Richardson.  You might
also look up more recent work by the LCR people.  Leroy Hood published
a Science paper in 1988 on the method, and R.B.Wallace has published
papers which specifically address your question.  You might want to
check out Taq ligase in order to give better discrimination.  Many
happy joins,

Brett Lindenbach
brett at borcim.wustl.edu
Dept of Molecular Microbiology
Washington University School of Medicine





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