Pilot Genomics Experiment
Calu-3 cell, Influenza A/VN/1203/04
- Seed Calu-3 cells passage 14, 1x10^6 cells/TC6 well.
4 well virus and 2 well mock/time point = 1 TC6 plate/time point
6 time point = 6 TC6 plate
- Change cell media.
- Wash cells with BSA-D12F twice.
- Inoculate VN1203 MOI=1
- Adsorption for 40 min at 37C. Tilt cells every 15 min.
- Remove virus and wash the cells with BSA-D12F once.
- Add 2ml of BSA-D12F with 0.5ug/ml of TPCK trypsin.
- Collect samples at 0, 1.5, 3, 7, and 12 hours post-adsorption.
- Collect samples at 24 hours post-adsorption.
- 2 well of VN1203 infected cells and 1 well of mock infected cells at each time-point.
- Remove media from the cells as much as possible.
- Rinse cells with 5mL cold 1X PBS.
- Lyse cells with 1mL Trizol reagent.
- Pipette up and down until cells have been dislodged and liquid is a uniform color and consistency.
- Transfer to 2mL tube and vortex thoroughly.
- Incubate 5 minutes at room temperature.
- Freeze at -80C until ready to ship.
Metadata is available in the sub-folders on the right.
Microarray data is also available.