- Select five individual colonies from bacteria plate. Circle the colonies and label them 1-5.
- Label five sterile culture tubes with the colony number, date, and initials.
- Using a 10 ml serological pipette, transfer 12 ml of LB into a sterile centrifuge tube.
- Using a p20 micropipette, transfer 12 μl of 25 mg/ml 25 mg/ml chloramphenicol and 12 μl of 50 mg/ml kanamycin.
- Shake the test tube to mix the contents.
- Using a serological pipette, transfer 2 ml of the mixture into each of the 5 culture tubes.
- Inoculate each culture tube with their respective colonies using an autoclaved toothpick or a sterile loop.
- Place the inoculated culture tubes in a shaking incubator.
- Turn on the incubator. Set the temperature to 37°C and the RPM to 220. Make sure that the hr setting is off.
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