can and cyh selection problems

Julie Glover GLOVER at rsbs.anu.edu.au
Sun Mar 26 19:39:57 EST 2000


Hi yeast people,

I have been having trouble using the can1 and cyh2 markers to select for
haploid cells after sporulation.  If I understand correctly, diploid strains
that are heterozygous for these markers should be sensitive to these drugs,
however I get diploid heterozygous strains (unsporulated) growing on levels
of cycloheximide 5 microgram/ml even though the haploid sensitive strains
don't grow at all on much lower concentrations.  Similarly diploids (hets)
grow on canavanine 120 micrograms/ml even when haploid sensitive strains
don't grow at 60 micrograms/ml.  I've been doing cyc selection experiments
on YPD plates and canavanine selection on arginine dropout plates.  I've
taken colones from these plates and shown that these are still diploid
cells.

Does anyone have any ideas - am I missing something obvious as I am quite
new to yeast work?

Thanks,
Julie


Dr Julie Glover

************************************************
Groupe Limagrain Pacific
Research School of Biological Sciences
The Australian National University
GPO Box 475 Canberra  ACT  2601
AUSTRALIA
Tel 61 26249 4549 
Fax 61 26279 8593
glover at rsbs.anu.edu.au
************************************************

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