For internal use
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m9
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LB (Luria Broth) 500mL
| Trypton | 5.0 g |
| Yeast Extract | 2.5 g |
| NaCl | 5.0 g |
LB Plate (solid) 300mL
| Trypton | 3.0 g |
| Yeast Extract | 1.5 g |
| NaCl | 3.0 g |
| Agar | 4.5 g |
Appropriate amount of antibiotics: 100 µg/mL Amp, 50 µg/mL Kan
M9 (15N, 1 L)
| Na2HPO4 | 12.8 g |
| KH2PO4 | 3.0g |
| NaCl | 0.5g |
| 15N-NH4Cl | 1g |
| Add 960mL H2O (or D2O for deuterated sample); adjust pH to 7.4 → Autoclave | |
| Dissolve 8g glucose in 20mL H2O → Filter | |
| 8mL 1M MgSO4, 2mL 0.1M CaCl2 → Autoclave | |
| 6mL 1mM FeSO4 → Filter | |
Note: alternative: NaH2PO4 (monobasic) = 4.75g, Na2HPO4 (dibasic) = 3.73g, KH2PO4 (monobasic) = 7.15g
M9 (15N, 1L, with vitamin)
| Na2HPO4 | 12.8g |
| KH2PO4 | 3.0g |
| NaCl | 0.5g |
| 15N-NH4Cl | 1g |
| Add 1L H2O; adjust pH to 7.4 → Autoclave | |
| Dissolve 8g glucose in 20mL H2O → Filter | |
| Before using it, add Amp, 8mL 1M MgSO4, 2mL 0.1M CaCl2, 6mL 1mM FeSO4, 1mL of 10mg/mL Thiamine, 1mL of 10mg/mL Biotin and all the glucose solution. | |
Nice practical guide on media for NMR isotope labeling
m9.txt · Last modified: 2018/03/12 04:10 by 127.0.0.1
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