IGEM:British Columbia/Protocols/Day/Overnight Culture

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Contents

Day/Overnight Cultures

Materials

  • Liquid LB Medium with appropriate antibiotic
  • Streak plate/glycerol stock/liquid culture with desired cells

Equipment

  • Wire loop/sterilized toothpicks/pipet +tips
  • Flame or biohazard hood
  • 14mL round-bottom tubes

Method

  • Add 1.5-5mL of LB broth to a round-bottom tube.
    • From glycerol stock: scrape the stock with a pipet tip and eject the tip into the broth. DO NOT thaw the stock.
    • From a colony: touch the colony with a toothpick/pipet tip and drop into the broth, or use a wire loop if you have one.
    • From liquid culture: pipet a few μL or use an inoculating loop.
  • Incubate at 37°C with shaking for not more than 18 hours. Make sure that the top is loose enough to allow for air exchange but not so loose that it will come off during incubation.
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