Taking Samples and Fluid Changing Flasks at the 3 Week Mark for Experimental DiNV Evolution in Dinn Cells
Cells were inoculated on the 26th of Sept see post here. There are 3 replicate flasks that have been left to grow and get infected in the 23C incubator for 21 days. I am only removing supernatant for samples, the cells are not getting passed throughout this experiment, the only thing that happens is a tri-weekly fluid change (once every three weeks).
All steps took place in the cell culture hood
- Warmed the 10% FBS, 4% mushroom Schneider’s medium with antibiotics in the hood to room temperature
- Prepared 1.5mL tubes to take samples
- Removed two samples 50ul from each flask before changing fluid and placed into the 1.5mL tubes:
| tube # | flask from |
|---|---|
| 11 | A |
| 12 | A |
| 13 | B |
| 14 | B |
| 15 | C |
| 16 | C |
- Removed 3mL of medium in each flask and discarded it into a 15mL conical. The conical was placed in the autoclave trash when full (should contain virus)
- Added 3mL of 10% FBS, 4% mushroom Schneider’s medium with antibiotics to each flask gently
- Gently rocked flasks to distribute the medium
- Placed flasks back in the 23C incubator for another 3 weeks
- Sample tubes were kept on ice, 3 were chosen for DNA extraction, the other 3 were frozen at -20
Sample information can be found here