Overview 

Pilot Genomics Experiment

Calu-3 cell, Influenza A/VN/1203/04  

Day 1

  1. 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

Day 2

  1. Change cell media.

Day 3

  1. Wash cells with BSA-D12F twice.
  2. Inoculate VN1203 MOI=1
  3. Adsorption for 40 min at 37C.  Tilt cells every 15 min.
  4. Remove virus and wash the cells with BSA-D12F once.
  5. Add 2ml of BSA-D12F with 0.5ug/ml of TPCK trypsin.
  6. Collect samples at 0, 1.5, 3, 7, and 12 hours post-adsorption. 

Day 4

  1. Collect samples at 24 hours post-adsorption.  

Harvesting Samples

  • 2 well of VN1203 infected cells and 1 well of mock infected cells at each time-point.
  • Procedure:
  1. Remove media from the cells as much as possible.
  2. Rinse cells with 5mL cold 1X PBS.
  3. Lyse cells with 1mL Trizol reagent. 
  4. Pipette up and down until cells have been dislodged and liquid is a uniform color and consistency.
  5. Transfer to 2mL tube and vortex thoroughly.
  6. Incubate 5 minutes at room temperature.
  7. Freeze at -80C until ready to ship.

 

Metadata is available in the sub-folders on the right.

Microarray data is also available.

 


Viral Titers 

See the Excel file attached below for data shown on this graph.

  Attached Files  
   
 image002.png
 ICL002viral titers.xls


Viral RNA 

See the attached Excel file for data.

  Attached Files  
   
 image002.png
 image004.png
 ICL002 qPCR data analysis released to Public.xls


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