Monday Jul. 27
Beginning of the manufacturing project! Let's do it!
Choice of the strain of yeast: Sc. M Cherry, wich is resistant to geneticin at a concentration of 200µL/mL
We will have to calculate a survival rate so we have to be able to know how many cells we have in the first place for each experiment:
Centrifuge the culture tube
Throw the media away
Replace it by a known quantity of osmosed water
Take the OD, doing the blank with osmosed water
Make dilutions of the yeasts/water solution to plate them
Deduce the concentration of Sc. M Cherry corresponding to 1 OD
First try today and plating of the different dilutions on YPD agar+ GEN (200)
Tuesday, Jul. 28
First idea: drying the yeasts. Protocol:
Centrifuge the culture tube
Throw the media away, replace it with a known quantity of osmosed water, vortex, take the OD
Centrifuge again, throw the water away keeping a very small amount of it to detach the yeasts from the tube easily
Spread on aluminium
Let it dry for 2 hours
Scratch the aluminium to detach the dried yeasts
Plate the powder obtained to see if Sc. M Cherry survived
Results:
The aluminium is not a good idea, the yeasts stick to it and it gets destroyed when we scratch. We have to find another drying paper. Wednesday, Jul. 29
Result of the first OD test:
The 10mL of solution had an OD of 2,557 and 100µL of the 10^-4 dilution grew 359 colonies.
so '''1 OD = 1,4x10^7 cells/mL'''
We launched a similar experience for more repetitions.
