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This is the media used for C. Hutch and co cultures with E. Coli. The media contains all the salts needed for both bacteria but not the carbon source (filter paper) due to the carbon source not being liquid. Cultures are made by adding the salt solution to a tube containing a piece of filter paper where the tube with the paper in it has been sterilized (usually by autoclave).
C Hutch ALONE in 1 L
Paper: 50 grams per liter of media, which is one 0.5g 9cm No. 1 filter paper disks per 100 mL.
| Component | Amount |
|---|---|
| KH2PO4 | 0.2 g |
| MgSO4*7H2O | 0.5 g |
| KCl | 0.5 g |
| NaNO3 | 0.5 g |
| FeSO4*7H2O | 20 mg |
| K2HPO4 | 0.8 g |
| tryptone | 1 g |
Autoclave 15 min.
E Coli ALONE (M9) in 1 L
- Make 5x M9 salts
- To make M9 Salts aliquot 800ml H2O and add
- 64g Na2HPO4-7H2O
- 15g KH2PO4
- 2.5g NaCl
- 5.0g NH4Cl
- Stir until dissolved
- Adjust to 1000ml with distilled H2O
- Sterilize by autoclaving
- To make M9 Salts aliquot 800ml H2O and add
- Measure ~700ml of distilled H2O (sterile)
- Add 200ml of M9 salts
- Add 2ml of 1M MgSO4 (sterile)
- Add 20 ml of 20% glucose (or other carbon source)
- Add 100ul of 1M CaCl2 (sterile)
- Adjust to 1000ml with distilled H2O
