Preparation of Overnight Cultures

  1. Obtain a 14 mL Sterile Polypropylene Round-Bottom Tube.
  2. Add ~ 3-5 mLs of an appropriate broth (either LB or TB)
  3. Add an appropriate antibiotic if necessary and set this tube safely aside
    • Amp (1:100)
    • Chlor (1:1000)
    • Kan (1:1000)
  4. Prick the desired colony off a plate and suspend it in 50 uL PBS.
    • Refer to the PBS stock making protocol!
  5. Add 10 uL of the PBS sample and add it to the broth previously set aside.
    • Incubate overnight cultures @ 37oC, overnight with shaking.