Is it OK to ligate at RT overnight?
ANDREI.POPOV at bbsrc.ac.uk
Tue Sep 19 18:17:42 EST 1995
> Blunt-end ligations are favored at lower (ie., 16 degrees) than at RT or
>higher, because the lower temperature decreases the kinetic energy of the
>ends so that ligase has time to do its job.
I would suggest just the opposite.
Cohesive ends have a chance to stick together at 16C, but
at higher temp. (e.g. 24C) blunt ends have greater chances to
meet each other. In fact I did blunt ligations for 1 hr at 37C
just for the fun of it and it worked very efficiently.
There was a paper in NAR a couple of years ago showing
that ligations in the ultrasonic bath go quicker
presumably because of the frenetic movements of the DNA ends.
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