In <49046a$fv8 at epervier.CC.UMontreal.CA>, Daniel Durocher <DurochD at IRCM.UMontreal.CA> writes:
> a demystification of the O/N ligation?
For cohesive ends, we routinely do our ligations for 3h at room temperature
in a reaction mixture that has up to 0.4% low-melt agarose (only because that's
where our fragments come from, usually). Apparently there is quite a bit of
leeway. I've never tried 5 minutes, though!