Using 12 Well Plates of Myd88 and Dv-1 Cells to Infect With DiNV and Take Day 0 Samples
Using the day 26 innubila primaries infected with DiNV fluid from Kent for this infection experiment.
Plate layouts:
| Day 0 | 1 | 2 | 3 | 4 | ||||
|---|---|---|---|---|---|---|---|---|
| A | Dv-1 Infected with DiNV replicate 1 | Myd88 Infected with DiNV replicate 1 | Dv-1 not infected replicate 1 | Myd88 not infected replicate 1 | ||||
| B | Dv-1 Infected with DiNV replicate 2 | Myd88 Infected with DiNV replicate 2 | Dv-1 not infected replicate 2 | Myd88 not infected replicate 2 | ||||
| C | Dv-1 Infected with DiNV replicate 3 | Myd88 Infected with DiNV replicate 3 | Dv-1 not infected replicate 3 | Myd88 not infected replicate 3 | ||||
| Day 5 | 1 | 2 | 3 | 4 | ||||
|---|---|---|---|---|---|---|---|---|
| A | Dv-1 Infected with DiNV replicate 1 | Myd88 Infected with DiNV replicate 1 | Dv-1 not infected replicate 1 | Myd88 not infected replicate 1 | ||||
| B | Dv-1 Infected with DiNV replicate 2 | Myd88 Infected with DiNV replicate 2 | Dv-1 not infected replicate 2 | Myd88 not infected replicate 2 | ||||
| C | Dv-1 Infected with DiNV replicate 3 | Myd88 Infected with DiNV replicate 3 | Dv-1 not infected replicate 3 | Myd88 not infected replicate 3 | ||||
- Thawed 6 40ul aliquots of the cell fluid with DiNV on ice
- Prepared 12 1.5mL tubes to put the supernatant of the day 0 wells into. Sample names can be seen here
- Did all processes in the cell culture hood
- Pipette mixed vials of DiNV fluid before using
- Added 20ul of DiNV fluid to wells A1, A2, B1, B2, C1, and C2 in both the day 0 and day 5 plates
- After adding to the well, I gently rocked the plate back and forth
- For the day 0 plate, removed all the supernatant from each well and put it in the respective 1.5mL tube
- Added 1mL of 1X PBS to each now “empty” well in the day 0 plate too have fluid for the cells
- Taped up the plate and put the supernatant tubes in a new box and put it all in the -80
- Put the day 5 plate in the 23C incubator until Monday