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HomeMammalian Cell CulturePowdered Media Preparation Instructions

Powdered Media Preparation Instructions

Preparation of Media for Filtration

Powdered media and salt mixtures are extremely hygroscopic and should be protected from atmospheric moisture. The entire contents of each package should be used immediately after opening. Preparing medium in concentrated form is not recommended as precipitates may form. Supplements can be added prior to filtration or introduced aseptically to sterile medium. The nature of the supplement may affect the storage conditions and shelf life of the medium.

Procedure

  • Measure out 90% of final volume of tissue culture grade water. Water should be at room temperature.
  • While gently stirring the water, add the powdered medium or salt mixture. Stir until dissolved. Do not heat water.
  • Rinse original package with a small amount of water to remove all traces of powder. Add to solution.
  • To the solution, add the required amount of sodium bicarbonate from the chart below for each liter of final volume of medium being prepared. Stir until dissolved.
  • While stirring, adjust the pH to 0.1-0.3 pH units below the desired pH since the pH may rise during vacuum filtration. The use of 1 N HCl or 1 N NaOH is recommended.
  • Bring medium to final volume with tissue culture grade water.
  • Sterilize immediately by filtration using a membrane with a porosity of 0.2 micrometers or less.
  • Aseptically dispense into sterile containers. Store liquid medium refrigerated at 2-8 °C and in the dark.
  • Osmolality will vary with the addition of sodium or potassium chloride, approximately 10 mOsm for 300 mg/L NaCl or 400 mg/L of KCl
Powdered MediaProduct No. NaHCO3
7.5% Solution (mL/L)
NaHCO3
Powder (g/L)
Ames' MediumA142025.71.932
BME EBSSB963829.32.2
DME/F-12 w/o phenol redD2906161.2
DME/F-12D9785161.2
DME/F-12D8900161.2
DME/F-12D0547161.2
DME HEPESD115249.33.7
DME w/o phenol redD290249.33.7
DME DeficientD503049.33.7
DMED552349.33.7
DMED564849.33.7
DMED777749.33.7
F-12 (Coon's Modification)F663635.72.676
Glasgow MEMG614836.72.75
HBSS w/o phenol redH13874.70.35
HBSSH23874.70.35
HBSS w/o phenol redH48914.70.35
HBSSH61364.70.35
IMDMI763340.33.024
KREBS-HENSELEITK3753282.1
KREBS-RINGERK400216.81.26
McCOY'S 5AM489229.32.2
MCDB-131M853715.71.176
MCDB-153M740315.71.176
M199 EBSS HEPESM252029.32.2
M199 EBSS w/o phenol redM376929.32.2
M199 EBSSM501729.32.2
M199 HBSSM03934.70.35
MEM EBSSM026829.32.2
MEM JOKLIKM051826.72
MEM EBSS NEAAM064329.32.2
MEM AlphaM064429.32.2
MEM AlphaM089429.32.2
MEM HBSS NEAAM10184.70.35
MEM EBSS HEPESM264529.32.2
MEM EBSS NEAA w/o phenol redM302429.32.2
MEM HBSSM46424.70.35
MEM EBSS AutomodM076929.32.2
NCTC 135N326229.32.2
Nutrient Mixture F-10N6635161.2
Nutrient Mixture F-12N676015.71.176
Nutrient Mixture F-12KN352033.32.5
RPMI-1640 DeficientR138326.72
RPMI-1640 HEPESR413026.72
RPMI-1640R650426.72
RPMI-1640 HybrimaxR800526.72
RPMI-1640 AutomodR775526.72
RPMI-1640 w/o phenol redR875526.72
Tyrode's SaltsT214513.31
Waymouth MB752/1W162529.92.24
Williams' Medium EW412529.32.2

Sodium Bicarbonate Amount
Materials
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