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Inoculating Liquid Culture on Plate Growth Medium

Items Required

  • Conical tube containing liquid culture (1 to 3 days of culture)
  • Plate growth medium
  • Spreader
  • Pipette (P20) and tip (P20)
  • Rubbing alcohol
  • Rubber gloves
  • Permanent marker
  • Kim towel
  • Garbage bag

Sample Procedure

  1. Wear rubber gloves and disinfect your hands with alcohol. Disinfect the clean bench with alcohol. Use alcohol to disinfect the items to be put in the clean bench.
  2. If water droplets have formed on the plate growth medium, knock off the water droplets on a Kim towel and let the medium dry for about 10 minutes.
  3. Hold the conical tube in your left hand (or right hand) and the pipette (20 μL) in your right hand (or left hand). Loosen the conical tube's cap, attach the tip to the pipette, and push it down lightly twice. *During this step, the tip must not touch anything. Since you are using both hands, the tip is prone to touch something. Be careful. If the tip touches something, replace it. Also be careful not to have the pipette touch the inside of the tube. Frequently wipe the outer surface of the pipette with alcohol.
  4. Open the conical tube's cap, use a pipette to extract the liquid culture, and gently place it on the center of the plate growth medium. *As a general rule, if the liquid culture has been cultured for 2 days, extract 20 μL to 40 μL. However, if you want to see colonies quickly or if the liquid culture has hardly changed and you still want to inoculate, inoculate 100 μL. *Hold the cap in your hand without putting it down. If you want to put it down, disinfect the surface with alcohol immediately before putting the cap on it. Or place the cap on a clean Kim towel.
  5. Put the cap on the conical tube and discard the tip.
  6. Put down the pipette and conical tube and take out the spreader from the packaging. *Carefully take out the spreader from the bag so it does not touch the bag's opening. If it touches the bag's opening, discard it.
  7. Hold the spreader with your right hand (or left hand) and set it on the plate growth medium. Then use your left hand (or right hand) to rotate the plate growth medium so that the culture is spread evenly over the entire plate. Then move the spreader three or four times vertically to imprint the culture onto the medium.
  8. Put on the cover, turn it over, and write the date, conical tube number, etc., on the edge.
  9. Set the incubator to 30°C and incubate.