Monday, May 31, 2010

Protocol for Preparation of RPMI-1640 Medium ( Complete ) from basal medium


Basal RPMI-1640 medium

Foetal Calf Serum

200mM L-Glutamine Sterile stock

Penicillin/Streptomycin 100x stock

500mL Duran-Schott bottle steriled


1. Thaw a bottle of Foetal Calf Serum ( FCS ) by placing it in 56 degree celcius water bath for 30 minutes.

2. Complete 400mL ( x10 ) of the RPMI-1640 medium by adding the components as follows:
  • Basal RPMI-1640 312mL
  • Foetal Calf Serum 80mL
  • 200mM L-glutamine strile stock 4mL
  • Penicillin/Streptomycin 100x stock 4mL

NOTE: Glutamine is unstable and should be kept frozen. Half-life in medium at 4 degree celcius is anout 3 weeks. Can be added to medium to be stored at -2o degree celcius ( i.e All complete medium with glutamine added to be stored at -20 degree celcius freezer ). Only the working medium bottle to be stored at 4 degree celcius for use.

Updated @ 12:58hrs

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