To make M1 medium:
Add the following to the 1L ddH2O, adjust to pH=7.4 and autoclave.
Component | Conc (mM) | MW (g/mol) | add per 1L (g) |
PIPES | 50 | 302.37 | 15.12 |
NaOH | 7.5 | 40 | 0.30 |
NH4Cl | 28 | 53.49 | 1.5 |
KCl | 1.3 | 74.55 | 0.10 |
NaH2PO4•H2O | 4.3 | 137.99 | 0.59 |
NaCl | 100 | 58.44 | 5.84 |
After autoclaving: | |||
100X vitamin stock | — | — | 10 mL |
100X mineral stock | — | — | 10 mL |
100 X amino acid stock | — | — | 10 mL |
Optional: | |||
Casamino acids | 0.2% | 2 g | |
sodium lactate | As needed |
100X vitamin stock:
Add the following to 1 L ddH2O, filter sterilize. Store at 4°C, protect from light.
vitamin | add to 1 L (mg) |
D-biotin (B7) | 2 |
folic acid (B9) | 2 |
pyridoxine HCl (B6) | 10 |
thiamine HCl (B1) | 5 |
nicotinic acid (B3) | 5 |
D-pantothenic acid, hemicalcium salt (B5) | 5 |
cobalamin (B12) | 0.1 |
p-aminobenzoic acid | 5 |
a-lipoic acid | 5 |
100X amino acid stock:
Add the following to 1 L ddH2O, adjust to pH=7.0. Filter sterilize and store at 4°C.
amino acid | add to 1 L (g) |
L-glutamic acid | 2 |
L-arginine | 2 |
D,L-serine | 2 |
100X trace mineral stock:
Add the following to 1 L ddH2O, adjust to pH=7.0. Filter sterilize and store at 4°C.
mineral | conc. (mM) | MW (g/mol) | add per 1 L (mg) |
C6H9NO6Na3 (Na-NTA) | 7.85 | 257 | 2017.4 |
MgSO4·7H2O | 12.17 | 256.47 | 3121.2 |
MnSO4·H2O | 2.96 | 169.02 | 500.3 |
NaCl | 17.11 | 58.44 | 1000.0 |
FeSO4·7H2O | 0.36 | 278.05 | 100.0 |
CaCl2·2H2O | 0.68 | 147.014 | 100.0 |
CoCl2·6H2O | 0.42 | 237.93 | 100.0 |
ZnCl2 | 0.95 | 136.315 | 129.5 |
CuSO4·5H2O | 0.040 | 249.7 | 10.0 |
AlK(SO4)2·12H2O | 0.021 | 474.39 | 10.0 |
H3BO3 | 0.016 | 61.83 | 1.0 |
Na2MoO4·2H2O | 0.010 | 241.95 | 2.4 |
NiCl2·6H2O | 0.010 | 237.69 | 2.4 |
Na2WO4·2H2O | 0.076 | 329.85 | 25.1 |