IGEM:Virginia 2012/Protocols/SSM Media Preparation
|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|
|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.
- 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).
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- Who has experience with this protocol?
or instead, discuss this protocol.