Arrange all the required items inside a sterile laminar flow hood designated for bacterial cell culture, or beneath a bunsen burner flame on an ethanol-sterilized work surface. Wipe down all tools with ethanol to maintain sterility.
Dispense 2 mL of medium into a culture tube. This makes a low-volume starting culture. To make larger cultures, scale up to higher volumes, e.g. dispense 5 mL of medium into a larger culture tube. Anything greater than this amount should be dispensed into culture flasks. Ensure that you do not put too much media in the culture vessel, since there must be enough space in it to allow air to circulate.
Pick a colony. Holding a sterile pipette tip attached to a pipettor, gently touch the chosen colony, being extremely careful not to come into contact with other colonies. If this happens, discard the tip and start again.
Inoculate the media. Open a culture tube with media and dip the tip of the pipette into the media. Swirl the tip or depress the plunger of the pipette to release or blow out the microbial colony. Cap the tube and place it into a tube rack.
Incubate overnight at 37 degrees Celsius. Secure the rack in the shaking incubator and set it to shake at 200 rpm, overnight (12-18 hours), at 37 degrees Celsius.