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

Add 3mL sterile/autoclaved LB in a 15mL Falcon tube.

Pipet 3μL of 1000X antibiotic into the LB.

Select a single colony using a sterile toothpick or pipette tip.

Place toothpick or pipette tip in the culture tube and stir.

Remove toothpick, or in the case of a pipette tip, leave in the tube.

Place culture tube in incubator at 37 degrees Celsius overnight shaking vigorously (250 RPM)