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Protocol 1 : Culture Medium preparation

Aim: Make bacteria population by culturing them on specific medium. Selecting the recombinant stain

Liquid LB medium preparation (1L)

Mix the following component in 1L of water :

  • 10g

  • 5g Yeast extract

  • 10g NaCl

    • Adjust the pH to 7
    Mix the solution until no lump left and add a magnetic stirrer
    Autoclave at 121 Celsius degrees for 15 to 30 minutes to sterilize
    Allow the autoclaved medium to cool to 55ºC (you can hold it in your hand) and add antibiotics (Chloramphenicol at a final concentration of 100 µg/mL.)

    Protocole for 30 plates of Luria-Bertani (LB) agar plates for growing E. Coli with standard plasmids (500mL)

    Mix the following component in 500mL of water:

  • 5g Tryptone

  • 2.5g Yeast extract

  • 5g NaCl

  • 7.5g Agar powder

    • Adjust the pH to 7.5
    Mix the solution until no lump left and add a magnetic stirrer
    Autoclave at 121 Celsius degrees for 15 to 30 minutes to sterilize
    Allow the autoclaved medium to cool to 55ºC (you can hold it in your hand) and add antibiotics (Chloramphenicol at a final concentration of 100 µg/mL).