ez007215 at rocky.ucdavis.edu (Zhaomin Yang) wrote:
> Now time has come for me to
>do some experiments with pombe. The problem I'm having is with the
>transformation of a ts pombe strain. I have tried a couple of times using
>the yeast transformation kit from ZYMO RESEARCH with no success.
>Wondering what protocols you guys usually use for pombe transformation.
>
We use either a modified LiOAc protocol, or electroporation. For
reference to the former, which came originally from Okayama's lab, see the Moreno et al.pombe review in the cerevisaie Methods Enzymol book (ed. Guthrie and Fink). For the EP, we use a similar protocol to the Guarente protocol for cerevisiae, except that 1) we grow pombe in selective media 2) we harvest them at mid log and 3) we don't plate them on sorbitol plates, but on regular selective plates.
I've tried a couple of those kits and I think they are optimised for cerevisaie; pombe doesnt seem to like them much.
susan
susan_forsburg at qm.salk.edu
