M4 Medium Recipe

To make M4 medium:                                                                                                                   

Add the following to the correct amount of distilled water

Component Conc (mM) MW (g/mol) add per 1L (g) add per 10L (g)
K2HPO4 1.27 174.18 0.221 2.21
KH2PO4 0.73 136.09 0.099 0.99
NaHCO3 2.00 84.01 0.168 1.68
(NH4)2SO4 9.00 132.14 1.189 11.89
NaCl 125.00 58.44 7.305 73.05
Hepes 5.00 238.3 1.192 11.92
Yeast extract 0.5 5.0
Tryptone 0.5 5.0
100X minerals 10 mL 100 mL
100X CaCl2 10 mL 100 mL
40% sodium lactate ADD AFTER

AUTOCLAVING!

4.3 mL 43 mL

 

For 100X Mineral Mix:

Add the following to 1 L distilled water. Store at 4°C.

component final mM g/mol final mg/L 100X g/L
Na2EDTA 0.0672 336.24 22.595328 2.260
MgSO4•7H2O 1.01 246.47 248.9347 24.89
MnSO4•4H2O 0.0013 223.07 0.289991 0.029
NaCl 0.01 58.44 0.5844 0.058
FeCl2 0.0054 126.751 0.6844554 0.068
CoCl2 0.005 129.839 0.649195 0.065
ZnSO4•7H2O 0.001 287.53 0.28753 0.029
CuSO4•5H2O 0.0002 249.7 0.04994 0.005
H3BO3 0.0566 61.83 3.499578 0.350
Na2MoO4 0.0039 205.92 0.803088 0.080
NiCl2•6H2O 0.005 237.69 1.18845 0.119
Na2SeO4 0.0015 188.94 0.28341 0.028

 

For 100X CaCl2 mixture:

Add 7.13 g CaCl2•2H2O to 1L H2O. Store at 4°C.

 

For 40% sodium lactate:

Dilute 60% lactate solution (as ordered) to 40% by pouring 333 mL 60% solution into a graduated cylinder and bringing the volume to 500 mL with distilled water. Mix well, filter sterilize and store at 4°C in the filter bottle.

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