( olume brought to 10 ml in SE...)Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae

olume brought to 10 ml in SE, and the beads incubated at 37 C for 2 to 24 hours. Following incubation, the beads were pelleted as before, resuspended in 20 ml of 25 mM Na₂EDTA pH 8.0, containing 1% sarkosyl (Na salt of N-lauroylsarcosine, Sigma) and 50 g/ml proteinase K (Boehringer Mannheim), and incubated overnight at 50 C. The next day they were centrifuged and washed several times in 20 ml aliquots of 50 to 150 mM EDTA, pH 7.5 to 8.0, before being stored as a slurry (approximately 10 ml) in the EDTA solution at 4 C for as long as 2 years.

The following substitutions⁹ have since produced identical results: SDS instead of sarkosyl; 1 mg/ml pronase E (Sigma P 6911) at 37 C for 2 to 24 hours instead of 50 g/ml proteinase K overnight at 50 C; stationary instead of log phase cultures; use of cells from individual colonies from a YEPD plate (0.25 ml agarose per colony); and a scaled-down but analogous cell work-up with microfuge tubes and 5-second microcentrifuge spins instead of 50 ml tubes and 5-minute low-speed spins.

Results
Electrophoretic Karyotype of the Strains
The karyotype (Figure 1) of the laboratory strains (lanes A, B, G, and H) is virtually identical to that previously described³, and hence the identities of the bands are also presumed to be the same. In the profile of the UCD 522A (lane C), band 1 migrates faster than the laboratory counterpart and although band 4 corresponds well to band 3 (chromosome III) in the laboratory strains, an additional faster migrating band is also present. The doublet corresponding to band 5 in the laboratory strains migrates faster, as does the UCD 522A version of laboratory band 7. Labo
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