See.my.signature wrote:
: Hi everyone
: I need to do some plasmid shuffle/gene replacement experiments, using the
: 5-FOA negative selection for loss of the URA3 marker. This stuff (FOA) is
: horrifically expensive! I have heard a rumour that there is a recipe for
: 5-FOA plates which enables one to use a lot less of the compound and get
: the same selection - does anyone out there have a ref for this or any info
: on the technique? post it or mail to me, and I will post a compilation of
: the answers for everyone.
: Thanks,
: colin macdiarmid
:macdiarm at sbsu1.auckland.ac.nz
the reference is McCusker, J. and Davis, R. _Yeast_ 7:607-608 (1991) The Use
of Proline as a Nitrogen Source causes Hypersensitivity to, and Allows more
Economical use of 5FOA in _Saccaromyces_cerevisiae_.
the technique doesn't work for all strain backgrounds so YMMV.
there is also a reference for a (relatively) inexpensive synthesis of FOA,
but I haven't looked this one up:
Alam, S.N.; Shires, T.K.; and Aboul-Eneinn, H.Y. _Acta Pharm. Suec._ 12:375
(1975).
--
Jeffrey Ziehler ziehler at post.its.mcw.edu
waiting for the clu- phenotype to be a target for gene therapy.
