Ultrasonic Ligation

the End jgraham at bronze.ucs.indiana.edu
Wed Aug 11 23:12:36 EST 1993


Max,

16C as a temperature for ligation represents a compromise of optimal 
temperature for annealing of short single stranded overhangs and that 
for ligase activity. Blunt end ligations are traditionaly done at 
room temperature as the first consideration is not important.

There does not appear to be a "routine ligation" as different combinations
of DNA fragments ligate with a widely different efficiencies. In approaching
a new combination of DNA fragments, I find it necessary to strive for the 
highest possible ligation efficiency as difficult constructs are
as common as theoretically "simple" ones. Several-fold differences 
in ligation efficiency can often make or break a plasmid construct
in my experience.

Jim
J. Graham
Biology and Chemistry Departments
Indiana Univeristy Bloomington



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