Maintenance ConA-fluorophore Label Under Standard Mating Conditions

The purpose of the experiment is to test if the ConA-fluorophore label is retained by the cells' surface under the standard mating conditions. Haploid-single-stained samples are obtained using optimized ConA-fluorophore conjugates concentrations, vacuum-filtered and incubated for 0.5, or 2.5 h in SD agar plates at 30ºC. Conclusion: By comparing the median ConA-Tet fluorescence intensity of the ConA-Tet positive subpopulations we determined that the older cell wall components (that is, mother cells' walls) retained more than 65 % of the initial fluorescent label after the mating incubation period. Unstained controls obtained at the different time points were included