Plating 2 24 Well Plates with Dinn-1 and Dv-1 Cells for DiNV Infection and qPCR
There will be 1 plate for the 0 hour and one plate for the 5 day timepoints.
Plate Layouts:
| 1 | 2 | 3 | 4 | 5 | 6 | |
|---|---|---|---|---|---|---|
| A | Dv-1 CC | Dinn CC | Dv-1 DiNV | Dinn DiNV | ||
| B | Dv-1 CC | Dinn CC | Dv-1 DiNV | Dinn DiNV | ||
| C | Dv-1 CC | Dinn CC | Dv-1 DiNV | Dinn DiNV | ||
| D | Dv-1 CC | Dinn CC | Dv-1 DiNV | Dinn DiNV |
Dinn Cells
- Cannot be counted because of clumpiness
- Scraped a flask and transferred solution to a 15mL conical
- Spun down the conical for 4 minutes at 800rpm
- Resuspended the cell pellet in 16mL fresh medium
- Added 1mL of cell solution to each planned Dinn well in the plates (see layout above)
Dv-1 cells
- Scraped a flask and transferred 3mL of solution to a 15mL conical
- Spun down the conical for 4 minutes at 800rpm
- Resuspended the cell pellet in 1mL fresh medium
- Used 20ul to count in a hemocytometer:
- Q1: 663
- Q2: 767
- Q3: 555
- Q4: 550
- Average: 634
- Cells per mL: 6,337,500
- Used 254ul of cells in 16mL of fresh medium
- Added 1mL of cell solution to each planned Dv-1 well in the plates (see layout above)
- This was about 100,000 cells per well
Placed plates in 23 incubator overnight