Cell Culture Preparation of SPZ Eggs
20211101 Making New Yeast Paste
- I made fresh yeast paste by using ~20mL of dry yeast, ~20mL of molecular grade water, and a few mL of apple juice. I added in some extra yeast to get the consistency right. Wanted it to be kind of like royal icing
- Autoclaved on program 3 once twice on 20211101
20211102 Making Yeast Plates
- Made sure the scooper/spatula was autoclaved
- In the afternoon, made 2 apple juice yeast plates with 2 streaks of yeast on each plate
- Made a new conical of 10% FBS medium:
- 5mL FBS serum
- 44.5mL 420 medium (the last of it!)
- 500ul 100X antibiotics
- 50ul gentaminicin
20211103 Setting up SPZ Flies to Lay
- Set up 4 vials of SPZ flies to lay on the yeast plate at about 2pm
- These flies were newly emerged within a week or so
20211104 Day 1 Cell Culture
- Collected the apple juice plate at ~9:45am and replaced the cage with a new plate for more laying
- Followed the Prepping Cell Culture Protocol
- Used 0% FBS medium made on 20211025 and 10% FBS medium made on 20211102
- Used Trypsin opened 20211028
- For added sterility: after every time doing something out of the TC hood, I sprayed my gloves with ethanol before going back into the hood (centrifuges, etc.)
- Flasks were left to settle overnight and checked the next day
- Supplies were put in the autoclave in the lab to sterilize for the next day
20211105 Day 2 Cell Culture
- Collected apple juice plate at ~9:30am, flies were placed in the morgue
- Followed the Prepping Cell Culture Protocol
- Used 0% FBS medium made on 20211025 and 10% FBS medium made on 20211102
- Used Trypsin opened 20211028
- For added sterility: after every time doing something out of the TC hood, I sprayed my gloves with ethanol before going back into the hood (centrifuges, etc.)
- Flasks were left to settle over the weekend before looking at them
Flasks were imaged on 20211108
Flasks made on 20211104 did not have that many cells in them:
Flasks made on 20211105 had a decent amount of cells in them, but I wouldn’t say a lot
