IGEM:Virginia 2012/Protocols/SSM Media Preparation: Difference between revisions
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| CaCl<sub>2</sub> 2H<sub>2</sub>O ''*20 mg/mL'' || 20mg || 40mg || 80mg || 100mg | | CaCl<sub>2</sub> 2H<sub>2</sub>O ''*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== | ==Procedure== | ||
Revision as of 11:32, 31 May 2012
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
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
References
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.
