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

8.1. Date#

24102022 (Monday)- 27102022 (Thursday)

8.2. Objective#

  • 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%

8.3. 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.

    • No dilution of cells .

8.4. Results#

  • Plate layout

Plate layout

  • Incubation in 30C at 12 pm

End of incubation curves

  • Measuring in 36C + 24h incubating in the biotek

End of measuring curves

  • Microscopy images

at 0% Gal WT,WT+pgal,dbem1+pgal,dbem1dbem3+pgal

at 0.01 Gal WT,WT+pgal,dbem1+pgal,dbem1dbem3+pgal

at 0.02 Gal WT,WT+pgal,dbem1+pgal,dbem1dbem3+pgal

at 0.05 Gal WT,WT+pgal,dbem1+pgal,dbem1dbem3+pgal