Procedure for the preparation of 20 liters medium

CCT_HK_IV_TD

Culture media Hektor, InVitrus, TurboDoma
Fortified culture media InVitrus VP-6, TurboDoma TP-6

  • Packaging
    Hektor, InVitrus & TurboDoma media kits consist of 5 vials containing liquid solutions.
    InVitrus VP-6 & TurboDoma TP-6 media kits consist of 6 vials containing liquid solutions.
    Sodium hydrogen carbonate and L-glutamine have to be added separately.
     
  • Storage
    Store the kit at -20ºC for optimal preservation.
     
  • Preparation
    After thawing, shake the vials vigorously before use!
    Time required for preparing the medium for sterile filtration: 10 minutes.
  •  


    1. Measure out 18 liters of double distilled or purified tissue culture water at ambient temperature
      in a stirred vessel. While stirring, add the solutions without heating the water and ensure full recovery of the material from the vials by rinsing them well.
       
    2. Add solutions no. 1 and no. 2, then exactly 45.0 g sodium hydrogen carbonate and wait until complete dissolution.
      The recommended L-glutamine concentration is 4mM (11.68 g for 20 liters medium).
      Note that higher glutamine levels may lead to excess cytotoxic catabolite formation.
       
    3. Hektor, InVitrus & TurboDoma media
      Add solutions no. 3, no. 4 and no. 5. Adjust end volume with water.
      Do not adjust pH (the media are buffered with MOPS)!

      InVitrus VP-6 & TurboDoma TP-6 media:
      Add solutions no. 3, no. 4, no. 5 and no. 6. Adjust both end volume and, if necessary, pH to 0.1-0.2 units below the desired value with gaseous carbon dioxide.  

    4. Sterilize by filtration and store refrigerated at 4-6ºC in the dark.

     


  • Warning
    The filter sterilized medium without L-glutamine expires 9 months after preparation.
    Do not store small volumes of culture media in large vessels and protect them from light!