Adding Dv-1 and Myd88 Cells to the 12 Well Plate for Transfection with GFP Plasmid, and Making 24 Well Plates for Myd88 Infection Assay

I was both adding cells to the plate that already had Dinn-1 cells for transfection with the GFP plasmid, as well as making plates for an infection test for the Myd88 cells. 1 12 well plate used for transfection. 2 24 well plates used for infection (a day 0 and a day 5).

Myd88 Cells

  • Passage 44 cells
  • Scrapped and pelleted, the cells were resuspended in 5mL of fresh medium
  • 20ul was counted in a hemocytometer:
    • Q1: 529
    • Q2: 555
    • Q3: 590
    • Q4: 482
    • Average: 539
  • Cells per mL = 5,390,000 cells
  • For the 12 well transfection plate:
    • 6mL of medium was needed
    • 150ul of cell solution was added
    • 1.5mL of solution was added to column 2 in the plate
    • This is ~200,000 cells added per well
  • For the 24 well infection plates:
    • 16mL of medium was needed
    • 600ul of cell solution was added
    • 1mL of solution was added to column 2 and column 5 in both plates (one for day 0 and one for day 5)
    • This is ~200,000 cells added per well
  • See layouts below for more info

Dv-1 Cells

  • Passage 83 cells
  • Scrapped and pelleted, the cells were resuspended in 11mL of fresh medium
  • 20ul was counted in a hemocytometer:
    • Q1: 430
    • Q2: 438
    • Q3: 364
    • Q4: 409
    • Average: 410
  • Cells per mL = 4,102,500 cells
  • For the 12 well transfection plate:
    • 6mL of medium was needed
    • 100ul of cell solution was added
    • 1.5mL of solution was added to column 1 in the plate
    • This is ~100,000 cells added per well
  • For the 24 well infection plates:
    • 16mL of medium was needed
    • 400ul of cell solution was added
    • 1mL of solution was added to column 2 and column 5 in both plates (one for day 0 and one for day 5)
    • This is ~100,000 cells added per well
  • See layouts below for more info

Layout for Transfection Plate:

  1   2   3   4  
A Dv-1 control   Myd88 control   Dinn 3 Control   Dinn 3 Control  
                 
                 
B Dv-1 transfected   Myd88 transfected   Dinn 3 transfected   Dinn 3 transfected  
                 
                 
C Dv-1 transfected   Myd88 transfected   Dinn 3 transfected   Dinn 3 transfected  
                 
                 

Layout for Infection Plate:

  1 2 3 4 5 6
A Dv-1 CC Myd88 CC X Dv-1 DiNV Myd88 DiNV X
B Dv-1 CC Myd88 CC X Dv-1 DiNV Myd88 DiNV X
C Dv-1 CC Myd88 CC X Dv-1 DiNV Myd88 DiNV X
D Dv-1 CC Myd88 CC X Dv-1 DiNV Myd88 DiNV X