IGEM:Virginia 2012/Protocols/SSM Media Preparation

Materials

BASAL MEDIUM to make	1 Liter	2 Liters	4 Liters	5 Liters
Glutamic acid, monosodium salt, 1H2O	10.72g	21.44g	42.88g	53.60g
Trisma Base	1.525g	3.05g	6.10g	7.63g
NaCl	2.50g	5.00g	10.00g	12.50g
KH2PO4	0.50g	1.00g	2.00g	2.50g
KCl *200 mg/mL	0.20g	0.40g	0.80g	1.00g
MgCl2 6H2O *100mg/mL (0.5 mM)	100mg	200mg	400mg	500mg
CaCl2 2H2O *20 mg/mL	20mg	40mg	80mg	100mg
Q.S. with deionized water to**	980mL	1960mL	3920mL	4900mL

Note: *Make stock solutions and add this amount. Note: ""These Volumes are to allow for 20 mL/L worth of supplements to be added later.

Note: For SSM-S, add 2.0 g/L of 2-ketoglutarate and pyruvate before pH'ing. Note: if pouring blood plates, bring volume up to 100 mL/L LESS than above. Note: it is convenient to make up the last three salts in a concentrated stock solution, such that you can use 1mL/liter of SSM.

Procedure

• Add 2.9g of NaCl to a 1 L bottle.
• Add in 25 mL of 7.5 pH Tris-Cl
• Bring this to a volume of 500 mL by adding distilled water.
• Autoclave (large liquid).

Notes

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