Team:GeorgiaTech/Agar Plates and Media Protocol


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  • Add (35 g/L of LB Agar) or (25 g/L LB broth and 15 g/L Agar) to a flask with distilled water
  • Dissolve and autoclave for 1 hour on fluid cycle
  • Antibiotics should be added at the correct concentration and volume when the agar has cooled to around 50°C or cool enough for hands to be touching the exterior of the flask.
  • Concentrations of antibiotics:
    • Chloramphenicol 34 µg/µL - Blue
    • Kanamycin 50 µg/µL - Green
    • Ampicillin 100 µg/µL - Red
  • For every mL of LB agar solution, add 1 µL of antibiotics.
    • Since I was using 300 mL of water, I added .3 mL of kanamycin
  • The plates are poured and cooled, then marked with a marker to signify the antibiotic.
  • Agar stabs are from the same protocol, just poured into tubes and capped.
  • Media solution is made from LB broth (25 g/L), autoclaved, and antibiotics added to it.
  • From the toothpick used to create an agar stab, the toothpick is placed into the media solution to start the culture.