Cell Culture Preparation of Dicer 2 Eggs

20211025 Prepping Apple Juice Plates

  • Made sure the spoon/scraper had been autoclaved
  • In the afternoon, made apple juice yeast plates with 2 streaks of yeast on each plate
  • Made fresh medium for this week:
  • 1 50mL conical of 10% serum media
    • 5mL of serum
    • 44.5mL of 420 Ex-Cell medium
    • 500ul of 100X antibiotics (Penicillin, Amphotericin B, and Streptomycin)
    • 50ul of 50mg/mL Gentamicin
  • 2 50mL conicals of media without serum
    • 49.5mL 420 Ex-Cell medium
    • 500ul of 100X antibiotics (Penicillin, Amphotericin B, and Streptomycin)
    • 50ul of 50mg/mL Gentamicin

20211026 Set Up Flies

  • Set up 3 vials of flies in the cage with the apple juice plate
  • This was around 3pm

20211027 Day 1 Cell Culture

  • Collected the apple juice plate at ~9:45am and replaced the cage with a new plate for more laying
  • Followed the protocol for preparing primary cell culture
  • Not very many eggs collected with these flies
  • Used 420 medium made on 20211025
  • For added sterility: after every time doing something out of the TC hood, I sprayed my gloves with ethanol before going back into the hood (centrifuges, etc.)
  • Flasks were left to settle overnight and checked the next day
  • Supplies were cleaned via protocol and autoclaved that day

20211028 Day 2 Cell Culture

  • Collected apple juice plate at ~9:45am, flies were placed in the morgue
  • Followed the protocol for preparing primary cell culture
  • More eggs than the day before, but not that many still
  • Used 420 medium made on 20211025
  • For added sterility: after every time doing something out of the TC hood, I sprayed my gloves with ethanol before going back into the hood (centrifuges, etc.)
  • Flasks were left to settle for a few hours before looking at them

The camera is being used in the fly room, so I did not take any good pictures. There are some single cells in the flasks, but not very many. This could be because of the poor laying of the flies.