Overnight Cultures

Reagents and Materials (per culture prepared):

  • 10mL culture tube. Use 16mm x 160mm or 16mm x 125mm
  • 5 mL LB
  • 5 uL 1000X antibiotics
  • Single colonies on a plate (best not to start an over night from a glycerol stock)


  1. Add 5mL sterile/autoclaved LB in a 10mL culture tube.
  2. Pipet 5μL of 1000X antibiotic into the LB.
  3. Select a single colony using a sterile toothpick or pipette tip.
  4. Place toothpick or pipette tip in the culture tube and stir.
  5. Remove toothpick, or in the case of a pipette tip, leave in the tube.
  6. Place culture tube in incubator at 37°C overnight shaking vigorously (250 RPM)