IGEM:Harvard/2010/General Protocols/EColi

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E. coli is a typical bacteria used for molecular cloning. Long-term storage of BioBrick plasmids is in E. coli suspended in glycerol (500ul cells:500ul glycerol) and kept at -80˚C. Growing E. coli is performed at 37˚C, at which temperature they divide every 20 minutes.

  • Day1: Making streaks from glycerol stocks
    • Warm agar plate at 37˚C for 10 minutes
    • Label lid of plate with what you're streaking on it, and your name, and the date
    • Locate desired glycerol stock in -80˚C freezer and, using a sterile toothpick, scrape out a tiny bit and streak on plate
    • Incubate plate overnite at 37˚C
  • Day2: Growing liquid cultures
    • Label 15mL falcon tube and add 5mL of LB media + selective antibiotic
    • Using 200uL tip w/o filter, touch streak or single colony and put tip in falcon tube
    • Grow culture overnite in a shaking 37˚C incubator
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