Originally published by Sager and Granick (1953) Ann. New York Acad. Sci. 56, 831-838
This recipe is a slight modification of their "medium I", as made in the laboratory of Dr. Joel Rosenbaum at Yale.
Make the following separate stock solutions:
1. trace elements
| H3BO3 | 1.0 g | |
| ZnSO4 . 7H2O | 1.0 g | |
| MnSO4 . 4H2O | 0.30 g | |
| CoCl2 . 6H2O | 0.20 g | |
| Na2MoO4 . 2H2O | 0.20 g | |
| CuSO4 | 0.04 g | |
| water to 1 liter |
2. sodium citrate
| Na citrate . 2H2O | 500 g | |
| water to 1 liter |
3. iron chloride
| FeCl3 . 6H2O | 10 g | |
| water to 1 liter |
4. calcium chloride
| CaCl2 . 2H2O | 53 g | |
| water to 1 liter |
5. magnesium sulfate
| MgSO4 . 7H2O | 300 g | |
| water to 1 liter |
6. ammonium salt
| NH4NO3 | 450 g | |
| or | ||
| NH4Cl | 600 g | |
| water to 1 liter |
|
Sager's original recipe specified ammonium nitrate; ammonium chloride should be substituted for strains lacking nitrate reductase activity
|
7. potassium phosphate, monobasic
| KH2PO4 . 7H2O | 200 g | |
| water to 1 liter |
8. potassium phosphate, dibasic
| K2HPO4 . 7H2O | 200 g | |
| water to 1 liter |
|
To make the final medium, mix 1.0 ml each of solutions 1 through 6, and 0.5 ml each of solutions 7 and 8.
The final pH should be about 6.9. To make nitrate medium for testing transformants of nit1, omit solution 6, and substitute 4.0 ml of a 1 M solution of KNO3. To grow non-photosynthetic strains, add 2.5 ml of a solution of 600 g/l sodium acetate hydrate (NaOAc . 3H2O), and increase solutions 7 and 8 to 1.0 ml each to increase the buffering capacity. |
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