Team:Ionis Paris/Protocol 1

Protocol 1 : Culture Medium preparation

Aim: Make bacteria population by culturing them on specific medium and/or selecting a recombinant strain

Liquid LB medium preparation (500mL)

Mix the following component in 500mL of water :

  • 5g Tryptone

  • 2.5g Yeast extract

  • 5g NaCl

    • Add a magnetic stirrer bar and mix the solution until there is no lump left
    Autoclave at 121 Celsius degrees for 15 to 30 minutes to sterilize
    Allow the autoclaved medium to cool to 55ºC (until you can hold it in your hand) and add antibiotics (Chloramphenicol at a final concentration of 25µg/mL.)

    LB Agar plates preparation (500mL)

    Mix the following component in 500mL of water:

  • 5g Tryptone

  • 2.5g Yeast extract

  • 5g NaCl

  • 7.5g Agar powder

    • Add a magnetic stirrer bar and mix the solution until no there is lump left
    Autoclave at 121 Celsius degrees for 15 to 30 minutes to sterilize
    Allow the autoclaved medium to cool to 55ºC (until you can hold it in your hand) and transfer it into a 50mL Falcon tube*
    Add 50µL antibiotics (Chloramphenicol at a final concentration of 25µg/mL) and mix*
    Poor 25mL of the liquid into an identified petri plate*
    Let the plate dry*

    *Steps to realize under the PSM class II

    Sources

  • Sigma-Aldrich - Instructions to microbial media