Difference between revisions of "IGEM:Virginia 2012/Protocols/SSM Media Preparation"
|Line 28:||Line 28:|
* to ..5-
* this .
Revision as of 10:35, 31 May 2012
|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.
- Adjust to pH 7.60 with concentrated HCl (about 0.5-0.7 mL/L).
- Autoclave for 20 min. (May store at this stage at 4°C as "SSM Incomplete.")'
- After cooling and before use, add 10 mL/L of both proline and SSM supplements (see below).
*Note: for SSM-S, also add 1 mL/L SSM-S supplement. May store basal medium at 4°C for short periods of time and at -20°C for somewhat longer.
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.