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

MATERIALS NEEDED:

Basal RPMI-1640 medium

Foetal Calf Serum

200mM L-Glutamine Sterile stock

Penicillin/Streptomycin 100x stock

500mL Duran-Schott bottle steriled

METHODS

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
• TOTAL VOLUME 400mL

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