Overnight Cultures

Reagents and Materials (per culture prepared):

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

Protocol

1. Add 3mL sterile/autoclaved LB in a 15mL Falcon tube.
2. Pipet 3μL 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 degrees Celsius overnight shaking vigorously (250 RPM)