Rapid Ligation Kit

Bruno Cenni cenni at cellbio.unige.ch
Thu Jan 6 06:10:34 EST 2000


You're right. The short ligation works well for simple (eg. up to 2-3 sticky end fragments
or 1 blunt insert) ligations.

"Frank O. Fackelmayer" wrote:

> As it is sometimes difficult to say
> if a particular cloning will be simple or not, I´d go for overnight ligation on a
> routine basis. It actually SAVES time in comparison to short ligations that don´t
> work   : )
>

Again, the transformation efficiency has often proven to be the crucial step in
problematic clonings...


Bruno


>
> Of course, if you are in a hurry and you may be happy with the short ligation.
> Remember, however, that not only ligation efficiency counts, but also transformation
> efficiency. So, use really good competent cells for all clonings (> 10e8 transformants
> per ug of supercoiled plasmid), especially when you cut down ligation time. I
> personally like electroporation because it never fails (well....)
>
> Frank





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