Pulsing Plates (Thymidine addition)

Always use radioactive tag to divert radiation

Keep everything sterile

Media with added thymidine will be in fridge in culture room in a yellow-capped tube.

  • IN HOOD: place white mat to absorb any liquid
  • Put plates on white mat in hood
  • Mix media and pour into small petri dish
  • Autopipette:
      • Located in bottom drawer next to hood on the left side
      • 10uL tips are located in white box on shelf next to hood on the right side
      • To load tip: Big lever down, small lever up. Slide in tip until it clicks. Push down small lever to lock
      • With thumb, push up big lever to fill the tip
      • Always push dispensing lever once to release air bubbles
      • To dispense, push lever once
      • To release tip, discard waste by pushing down big lever, lift up small lever and slide out tip. Replace tip into packaging
  • Dispense 10uL media into each well with media already in
  • Once added, replace plates in numerical order in the incubator (put radioactive tape on top plate)

 

Notes:

  • If there is not enough media, fill wells from top to bottom starting from the right side as those wells are the most important.
  • Use a 1mL pipette to get out any bits of media in yellow tube
  • Discard left over media back into original yellow-capped tube
  • Put used tip(s) in a 50mL waste tube.
  • Wrap up all thymidine waste in the white mat and tape up with radioactive tape. Place parcel in white icebox on windowsill.