Minimal (-trp) medium for bacteria
Prepare the following autoclaved stocks:
| i. | 20% (w/v) magnesium sulfate |
| ii. | 4 mg/ml uracil |
| iii. | 4 mg/ml histidine |
| iv. | 4 mg/ml leucine |
| v. | 20% (w/v) glucose |
Prepare the following filter-sterilized solutions:
| vi. | 50 mg/ml ampicillin |
| vii. | 1% thiamine hydrochloride |
Autoclave the following two solutions separately:
| viii. | 15 g of agar in 800 ml of distilled H2O |
| ix. | 10.5 g of potassium phosphate (dibasic) 4.5 g of potassium phosphate (monobasic) 1 g of ammonium sulfate 0.5 g of sodium citrate 160 ml of distilled H2O |
Cool Solutions viii and ix to 50°C, and mix them together. Quickly add 1 ml of Solution i, 10 ml of Solution ii, 10 ml of Solution iii, 10 ml of Solution iv, 10 ml of Solution v, 1 ml of Solution vi, and 0.5 ml of Solution vii. Mix the final solution well and pour the plates immediately.
