Plating Dinn-1 Cells for Transfection with GFP Plasmid

  • Using a 12 well plate
  • All processes took place in the cell culture hood
  • Took a confluent flask of cells and cell scraped it
  • Removed fluid from flask to 15mL conical
  • Centrifuged conical for 5 minutes at 200rpm
  • Discarded supenatant
  • Resuspended the pellet in 1mL of fresh medium
  • Passed the 1mL of cell mixture through at 22 gauge needle and syringe 20 times
  • Increased medium volume to 10mL
  • Added 1.5mL of cell mixture to the last two columns of wells in the 12 well plate
  • Placed plate in the 23C incubator for a few days for the cells to grow