DAS Vitamin Cocktail

To approximately 85 mL of dH₂O add Tricine while stirring continuously.
Adjust the pH to 8.0.
Add each of the remaining components in the order listed.
Bring total volume to 100 mL with dH₂O.
Sterilize the vitamin cocktail by filtration through a 0.45 µm Millipore filter.
Store in the dark at freezer temperature.

#	Component	Amount	Stock Solution Concentration	Final Concentration in DAS
1	Tricine (adjust to pH 8) (Sigma T 5816-256)	0.9 g/100 mL
2	Thiamine (Sigma T 1270)	0.067 g/100 mL	2.0 mM	0.99 µM
3	Biotin (Sigma B 4639)	0.001 g/100 mL	0.041 mM	0.02 µM
4	Vitamin B₁₂ (add after autoclaving) (Sigma V 6629)	0.0008 g/100 mL	0.0059 mM	0.0029 µM
5	Pantothenate (Sigma P 2250)	0.002 g/100 mL	0.091 mM	0.046 µM
6	PABA (p-aminobenzoic acid) (Sigma A 1174)	0.004 g/100 mL	0.291 mM	0.15 µM