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

Sodium Bicarbonate Amount

Powdered Media	Product No.	NaHCO3 7.5% Solution (mL/L)	NaHCO3 Powder (g/L)
Ames' Medium	A1420	25.7	1.932
BME EBSS	B9638	29.3	2.2
DME/F-12 w/o phenol red	D2906	16	1.2
DME/F-12	D9785	16	1.2
DME/F-12	D8900	16	1.2
DME/F-12	D0547	16	1.2
DME HEPES	D1152	49.3	3.7
DME w/o phenol red	D2902	49.3	3.7
DME Deficient	D5030	49.3	3.7
DME	D5523	49.3	3.7
DME	D5648	49.3	3.7
DME	D7777	49.3	3.7
F-12 (Coon's Modification)	F6636	35.7	2.676
Glasgow MEM	G6148	36.7	2.75
HBSS w/o phenol red	H1387	4.7	0.35
HBSS	H2387	4.7	0.35
HBSS w/o phenol red	H4891	4.7	0.35
HBSS	H6136	4.7	0.35
IMDM	I7633	40.3	3.024
KREBS-HENSELEIT	K3753	28	2.1
KREBS-RINGER	K4002	16.8	1.26
McCOY'S 5A	M4892	29.3	2.2
MCDB-131	M8537	15.7	1.176
MCDB-153	M7403	15.7	1.176
M199 EBSS HEPES	M2520	29.3	2.2
M199 EBSS w/o phenol red	M3769	29.3	2.2
M199 EBSS	M5017	29.3	2.2
M199 HBSS	M0393	4.7	0.35
MEM EBSS	M0268	29.3	2.2
MEM JOKLIK	M0518	26.7	2
MEM EBSS NEAA	M0643	29.3	2.2
MEM Alpha	M0644	29.3	2.2
MEM Alpha	M0894	29.3	2.2
MEM HBSS NEAA	M1018	4.7	0.35
MEM EBSS HEPES	M2645	29.3	2.2
MEM EBSS NEAA w/o phenol red	M3024	29.3	2.2
MEM HBSS	M4642	4.7	0.35
MEM EBSS Automod	M0769	29.3	2.2
NCTC 135	N3262	29.3	2.2
Nutrient Mixture F-10	N6635	16	1.2
Nutrient Mixture F-12	N6760	15.7	1.176
Nutrient Mixture F-12K	N3520	33.3	2.5
RPMI-1640 Deficient	R1383	26.7	2
RPMI-1640 HEPES	R4130	26.7	2
RPMI-1640	R6504	26.7	2
RPMI-1640 Hybrimax	R8005	26.7	2
RPMI-1640 Automod	R7755	26.7	2
RPMI-1640 w/o phenol red	R8755	26.7	2
Tyrode's Salts	T2145	13.3	1
Waymouth MB752/1	W1625	29.9	2.24
Williams' Medium E	W4125	29.3	2.2