6. Title : how do yeast cells look like during log phase at different galactose concentrations? (2nd experiment)#

6.1. Date#

07112022 (Monday)- 11112022 (Thursday)

6.2. Objective#

  • Take images with a proper density because the previous experiment with the original densities was too crowded for the WT strain.

  • Show that during log phase yeast cells with the gal promoter look OK at different galactose concentrations.

  • Genotypes:

    • WT+sfGFP+pGal-CDC42(ywkd065a)

    • dbem1+sfGFP+pGal-CDC42(ywkd069b)

    • dbem1dbem3+sfGFP+pGal-CDC42(ywkd070a)

    • WT(yll3a Control)

  • Gal concentrations:

    • 0%, 0.01%,0.02%,0.05%,0.06%,0.08%,0.1%,0.2%,2%

6.3. Method#

  • Population growth measurements in 0,0.01,0.02,0.05,0.06,0.08,0.1,0.2,2% galactose for the following strains: WT+sfGFP+pGal-CDC42(ywkd065a),dbem1+sfGFP+pGal-CDC42(ywkd069b),dbem1dbem3+sfGFP+pGal-CDC42(ywkd070a),WT(yll3a Control).

  • In the log phase of the measuring part take some bright field images of the cell morphology in the 96 well plate.

    • I transfer 50ul of cells to an 96 well imaging plate to take the bright field images.

    • Dilution of cells:

      • WT: all galactose concentrations: 1:1000

      • WT+pGal : more than 0.02% gal: 1:100

      • dbem1+pGal : more than 0.06% gal: 1:100

      • dbem1dbem3+pGal : more than 0.05% gal: 1:100

6.4. Results#