Is it OK to ligate at RT overnight?

Andrei Popov 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.

regards

Andrei Popov




More information about the Methods mailing list