Culture Media Preparation Technique

Posted on Mar 12, 2025 in Geology

Culture Media Preparation

According to the presentation: Dew refers to single-phase (solid, semi-solid, and liquid) and two-phase media.

General Technique for Preparing Culture Media

  1. Weighing: Carefully weigh the extract components or commercial dry media on a watch glass or in a beaker (depending on the quantity).
  2. Hydration: Add the required volume of distilled water.
  3. Dissolving: Stir the mixture in distilled water and heat to boiling. Maintain boiling for 1 to 2 minutes.
  4. pH Adjustment: Check the pH and adjust if there is any variation.
  5. Sterilization Container: Pour the media into a container for sterilization.
  6. Aseptic Distribution:
    • If the media was sterilized in a large container, distribute it aseptically into individual containers.
    • If distributing onto plates, place them near a burner to flame the mouth of the container before and after pouring.
    • Cover quickly and rotate to homogenize the distribution.
    • Keep plates uncovered for the shortest time possible to prevent desiccation and contamination.
    • Be careful not to pour the liquid when it is too hot. If necessary, perform a second, gentler sterilization (shorter time and lower temperature) in tubes.
    • The amount of media deposited in each container depends on its intended use.
  7. Cooling: Cool to ambient temperature before placing it in the refrigerator. If preparing agar slant tubes, allow them to solidify in a slanted position.
  8. Labeling: Label the batch.
  9. Storage: Store in the refrigerator or a cool place, covered, to prevent desiccation (especially plates).
  10. Ambient Temperature Equilibration: Equilibrate to laboratory temperature before use. Remove the media from storage with sufficient time before inoculation.
  11. Drying: Dry out if there is any condensation.

Culture Media Preparation

According to the presentation: Dew refers to single-phase (solid, semi-solid, and liquid) and two-phase media.

General Technique for Preparing Culture Media

  1. Weighing: Carefully weigh the extract components or commercial dry media on a watch glass or in a beaker (depending on the quantity).
  2. Hydration: Add the required volume of distilled water.
  3. Dissolving: Stir the mixture in distilled water and heat to boiling. Maintain boiling for 1 to 2 minutes.
  4. pH Adjustment: Check the pH and adjust if there is any variation.
  5. Sterilization Container: Pour the media into a container for sterilization.
  6. Aseptic Distribution:
    • If the media was sterilized in a large container, distribute it aseptically into individual containers.
    • If distributing onto plates, place them near a burner to flame the mouth of the container before and after pouring.
    • Cover quickly and rotate to homogenize the distribution.
    • Keep plates uncovered for the shortest time possible to prevent desiccation and contamination.
    • Be careful not to pour the liquid when it is too hot. If necessary, perform a second, gentler sterilization (shorter time and lower temperature) in tubes.
    • The amount of media deposited in each container depends on its intended use.
  7. Cooling: Cool to ambient temperature before placing in the refrigerator. If preparing agar slant tubes, allow to solidify before.
  8. Labeling: Label the batch.
  9. Storage: Keep in the refrigerator or a cool place, covered to prevent desiccation (mostly plates).
  10. Ambient Temperature Equilibration: Equilibrate to laboratory temperature before use. Remove them with sufficient notice before the inoculation.
  11. Drying: Dry out if there is water condensation.