Cell Culture Preparation of Domeless Eggs
20211109 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 0% FBS medium:
- 44.5mL 420 medium opened this day
- 500ul 100X antibiotics
- 50ul gentaminicin
20211110 Setting up Domeless Flies to Lay
- Set up 4 vials of Domeless flies to lay on the yeast plate at about 2pm
20211111 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 20211109 and 10% FBS medium made on 20211102
- Used Trypsin opened 20211111
- 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.)
- Yield of eggs was very good
- They made a nice egg pellet
- After processing, the cell pellet was smaller
- And there was a small issue with the homogenized cells clumping with each other in the tube and in the flask. I tried pipette mixing many times to break it up and have the flasks be even
- 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
20211112 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 20211109 and 10% FBS medium made on 20211102
- Used Trypsin opened 20211111
- 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.)
- Again there was a lot of eggs, even more than the day before I thought
- And they made a large egg pellet
- Flasks were left to settle over the weekend before looking at them
Flasks were imaged on the following Monday
Flask from 20211111:
Flask from 20211112
