ligation reaction

[Tom Knight]

Duncan Clark <blackhole at abuse.plus.com> writes:
> >Do you heat kill you ligation mix?
> 
> For electroporation yes because many years ago (I have been around
> that long!) it was reported that the T4 DNA ligase really lowers the
> electroporation efficiency. Heat killing that was the key to fixing
> the problem.

While this is probably true for normal ligation buffers, NEB
recommends against heat killing ligase in the quick ligation buffers.
Our experiments confirm about a 100x reduction in
ligation/transformation efficiency due to heat killing ligase in this
buffer.