Doris Germain (germain at PETERMAC.UNIMELB.EDU.AU) wrote:
: I have a protocol for MMS mutagenesis for pombe which use 2% MMS
: for 3 hours at room temperature, this treatment kill completely cerevisiae,
: does someone know what concentration would be more appropriate?
: Thank in advance
: Doris Germain
: Trescowthick Research Laboratories
: Peter MacCallum Cancer Institute
: Locked Bag 1
: A'Beckett Street
: Melbourne VIC 3000
In our recombination expt's we use concs of MMS from ~50 to 300 microg/ml
overnight. This is for a diploid strain, 300 microg/ml gives ~ 1% survival
after o/n, there may be strain differences and haploids will be much more
sensitive (300 microg/ml -> <0.1% surv.). Also, you will need higher concs
due to the shorter exposure time. I'd suggest doing a quick survival curve
for your strain between 50 and 500 microg/ml to get the correct % survival.
For a rough estimate about 200 microg/ml (0.2%) should do it for a haploid.
Hope this helps
brennan at mbcrr.harvard.edu