M1 Medium

To make M1 medium:                                                                                                                   

Add the following to the 1L ddHO, 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

 

 

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