DMEM

Medium for 3t3 fibroblasts and B16 cells

To make	mL DMEM
start with	mL water
add:	g DMEM*
add:	g NaHCO3
add:	g HEPES, then pH to 7.2 (initial pH = ~7.1)
add:	mL FBS
add:	mL anti/anti

bring to final volume, filter sterilize in BSC, refrigerate

• if using DMEM without pyruvate, add 1 mL Na Pyruvate (100mM) per 100 mL medium

Ingredient	supplier	cat #	1 L	500 mL	250 mL
DMEM			13.4	6.7	3.35	g
NaHCO3			3.7	1.85	0.925	g
HEPES			1.3	0.65	0.325	g
FBS	Krackeler	12103C	100	50	25	mL
anti/anti	Fisher	SV30079.01	10	5	2.5	mL

Mix DMEM, NaHCO3, HEPES in about 70% of the final volume of dH2O.
Initial pH ~7.5
Adjust pH to 7.2 with HCl (it will rise to 7.3 in the CO2 incubator).
Add appropriate aliquot of FBS and antibiotic/antimycotic.
Bring to final volume.
Filter sterilize in the tissue culture hood.
Store in the refrigerator.