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The medium composition is currently in its beta status, meaning that some concentrations are not correct and need further adjustments. Keep this in mind while trying this out.

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Current criteria: Folic acid

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11a MODIFIED MEDIUM 11

Compound Conc. [g/L] Conc. [mM]
Glucose 19.8 109.902
Meat extract 9.9 -
Casein peptone 9.9 -
Na-acetate 4.95 60.341
Yeast extract 4.95 -
K2HPO4 1.98 11.368
(NH4)3 citrate 1.98 8.141
Tween 80 0.99 0.756
MgSO4 x 7 H2O 0.198 0.803
MnSO4 x H2O 0.05 0.293
Pyridoxine hydrochloride 1.0e-4 4.9e-4
Nicotinic acid 5.0e-5 4.1e-4
Thiamine-HCl x 2 H2O 5.0e-5 1.3e-4
Riboflavin 5.0e-5 1.3e-4
Calcium pantothenate 5.0e-5 1.0e-4
Biotin 2.0e-5 8.2e-5
Folic acid 2.0e-5 4.5e-5
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Solutions:
  • 10 ml Vitamin solution

78a CHOPPED MEAT MEDIUM FOR TREPONEMA SP.

Compound Conc. [g/L] Conc. [mM]
Ground beef 473.934 -
Casitone 28.436 -
Agar 14.218 42.274
K2HPO4 4.739 27.21
Yeast extract 4.739 -
L-Glutamine 0.664 4.54
L-Histidine 0.569 3.666
L-Serine 0.474 4.51
L-Cysteine HCl 0.474 3.007
Ethanol 0.18 3.909
Vitamin K3 0.047 -
Hemin 0.005 0.007
Resazurin 9.5e-4 0.004
p-Aminobenzoic acid 2.4e-4 0.002
alpha-lipoic acid 2.4e-4 0.001
Pantothenic acid 2.4e-4 0.001
Thiamine-HCl x 2 H2O 2.4e-4 6.3e-4
Riboflavin 2.4e-4 6.3e-4
Vitamin B12 2.4e-4 1.7e-4
Nicotine amide 1.2e-4 9.7e-4
Nicotinic acid 1.2e-4 9.6e-4
Biotin 9.5e-5 3.9e-4
Folic acid 9.5e-5 2.1e-4
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Solutions:
  • 10 ml Haemin solution
  • 10 ml Vitamin K3 solution
  • 10 ml Vitamin K1 solution
  • 5 ml Vitamin solution
  • 50 ml Amino acid solution (filter sterilized)

78c CHOPPED MEAT MEDIUM FOR FRETIBACTERIUM SP.

Compound Conc. [g/L] Conc. [mM]
Ground beef 586.166 -
Casein peptone 35.17 -
Na-formate 7.034 103.43
Na-fumarate 7.034 43.953
K2HPO4 5.862 33.654
Yeast extract 5.862 -
Ethanol 2.227 48.349
L-Cysteine HCl x H2O 0.586 3.337
DL-Dithiothreitol 0.352 2.279
Haemin 0.003 0.004
Sodium resazurin 5.9e-4 0.002
Pyridoxine hydrochloride 1.2e-4 5.7e-4
Nicotinic acid 5.9e-5 4.8e-4
p-Aminobenzoic acid 5.9e-5 4.3e-4
(DL)-alpha-Lipoic acid 5.9e-5 2.8e-4
Thiamine HCl 5.9e-5 1.7e-4
Riboflavin 5.9e-5 1.6e-4
Calcium D-(+)-pantothenate 5.9e-5 -
Biotin 2.3e-5 9.6e-5
Folic acid 2.3e-5 5.3e-5
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Solutions:
  • 50 ml Clarified rumen fluid
  • 20 ml Fatty acid mixture
  • 1 ml Wolin's vitamin solution (10x)
  • 5 ml Haemin solution
  • 2 ml Vitamin K1 solution
Gas phase:
  • 80 % N2
  • 20 % CO2

78d MEDIUM FOR TREPONEMA PARVUM

Compound Conc. [g/L] Conc. [mM]
Ground beef 500 -
Casitone 30 -
Agar 15 44.599
K2HPO4 5 28.707
Glutathione 1.4 4.556
Na-pyruvate 0.5 4.544
L-Cysteine HCl 0.5 3.172
N-Acetylglucosamine 0.5 2.26
Ethanol 0.19 4.124
Casamino acids 0.16 -
Yeast extract 0.16 -
Tryptone peptone 0.16 -
Ascorbic acid 0.1 0.568
L-Glutamine 0.07 0.479
L-Histidin 0.06 0.387
L-Serin 0.05 0.476
Vitamin K3 0.05 -
Resazurin 1.0e-3 0.004
Hemin 5.0e-4 7.7e-4
p-Aminobenzoic acid 2.5e-4 0.002
alpha-lipoic acid 2.5e-4 0.001
Pantothenic acid 2.5e-4 0.001
Thiamine-HCl x 2 H2O 2.5e-4 6.7e-4
Riboflavin 2.5e-4 6.6e-4
Vitamin B12 2.5e-4 1.8e-4
Nicotine amide 1.3e-4 0.001
Nicotinic acid 1.3e-4 0.001
Biotin 1.0e-4 4.1e-4
Folic acid 1.0e-4 2.3e-4
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Solutions:
  • 10 ml Haemin solution
  • 10 ml Vitamin K3 solution
  • 10 ml Vitamin K1 solution
  • 5 ml Vitamin solution
  • 5 ml Amino acid solution
  • 5 ml Peptone mixture
  • 5 ml Hemin solution
  • 5 ml Ascorbic acid solution 2%(w/v)
  • 5 ml N-acetylglucosamin- and pyruvate solution
  • 10 ml Glutathione solution 14%(w/v)
  • 10 ml Sugar solution
Gas phase:
  • 80 % N2
  • 20 % CO2

81 MINERAL MEDIUM FOR CHEMOLITHOTROPHIC GROWTH (H-3)

Compound Conc. [g/L] Conc. [mM]
Agar 20.101 59.764
Na2HPO4 x 2 H2O 2.915 20.531
KH2PO4 2.312 16.986
NH4Cl 1.005 18.789
MgSO4 x 7 H2O 0.503 2.039
Ferric ammonium citrate 0.05 0.192
CaCl2 x 2 H2O 0.01 0.068
NaVO3 x H2O 0.005 0.036
Nicotinic acid 0.003 0.02
Pyridoxine hydrochloride 0.003 0.012
Calcium pantothenate 0.003 0.005
Thiamine-HCl x 2 H2O 0.003 -
H3BO3 0.002 0.024
CoCl2 x 6 H2O 0.001 0.008
ZnSO4 x 7 H2O 5.0e-4 0.002
Riboflavin 5.0e-4 0.001
Na2MoO4 x 2 H2O 1.5e-4 6.2e-4
MnCl2 x 4 H2O 1.5e-4 -
NiCl2 x 6 H2O 1.0e-4 7.8e-4
CuCl2 x 2 H2O 5.0e-5 2.9e-4
Vitamin B12 5.0e-5 3.7e-5
Folic acid 1.0e-5 2.3e-5
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Solutions:
  • 5 ml Standard vitamin solution
  • 50 ml Solution A
  • 920 ml Solution B
  • 20 ml Solution C
  • 5 ml Trace element solution SL-6

87a CHLOROFLEXUS AGGREGANS MEDIUM

Compound Conc. [g/L] Conc. [mM]
Glycyl-glycine 0.952 -
Yeast extract 0.952 -
NaNO3 0.476 5.603
NaCl 0.095 1.63
KNO3 0.095 0.942
Na2HPO4 x 2 H2O 0.095 0.671
MgSO4 x 7 H2O 0.095 0.386
CaCl2 x 2 H2O 0.048 0.324
Na3-EDTA 0.002 -
Nicotinic acid 9.5e-4 0.008
Thiamine-HCl x 2 H2O 9.5e-4 -
p-Aminobenzoic acid 4.8e-4 0.003
Pyridoxine hydrochloride 4.8e-4 0.002
Folic acid 4.8e-4 0.001
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Solutions:
  • 1 ml Trace element solution SL-6
  • 1 ml Vitamin solution CA

92a MODIFIED MEDIUM 92

Compound Conc. [g/L] Conc. [mM]
Trypticase soy broth 29.7 -
Agar 14.85 44.153
Yeast extract 2.97 -
Pyridoxine hydrochloride 1.0e-4 4.9e-4
Nicotinic acid 5.0e-5 4.1e-4
Thiamine-HCl x 2 H2O 5.0e-5 1.3e-4
Riboflavin 5.0e-5 1.3e-4
Calcium pantothenate 5.0e-5 1.0e-4
Biotin 2.0e-5 8.2e-5
Folic acid 2.0e-5 4.5e-5
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Solutions:
  • 10 ml Vitamin solution

113a SULFURIMONAS HONGKONGENSIS MEDIUM

Compound Conc. [g/L] Conc. [mM]
NaCl 24.9 426.085
Na2S2O3 x 5 H2O 4.98 20.066
KNO3 1.992 19.703
KH2PO4 1.992 14.638
Na2CO3 0.996 9.397
NH4Cl 0.996 18.621
MgSO4 x 7 H2O 0.797 3.233
FeSO4 x 7 H2O 0.002 0.007
Na2-EDTA 1.0e-3 0.003
H3BO3 6.0e-4 0.01
CoCl2 x 6 H2O 4.0e-4 0.003
ZnSO4 x 7 H2O 2.0e-4 6.9e-4
Pyridoxine hydrochloride 1.0e-4 4.8e-4
MnCl2 x 4 H2O 6.0e-5 3.0e-4
Na2MoO4 x 2 H2O 6.0e-5 2.5e-4
Nicotinic acid 5.0e-5 4.0e-4
p-Aminobenzoic acid 5.0e-5 3.6e-4
(DL)-alpha-Lipoic acid 5.0e-5 2.4e-4
Calcium D-(+)-pantothenate 5.0e-5 2.1e-4
Thiamine HCl 5.0e-5 1.5e-4
Riboflavin 5.0e-5 1.3e-4
NiCl2 x 6 H2O 4.0e-5 3.1e-4
CuCl2 x 2 H2O 2.0e-5 1.2e-4
Biotin 2.0e-5 8.2e-5
Folic acid 2.0e-5 4.5e-5
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Solutions:
  • 942 ml Solution A
  • 40 ml Solution B
  • 20 ml Solution C
  • 1 ml Solution D
  • 1 ml Solution E
  • 2 ml Trace element solution SL-4
  • 1 ml Wolin's vitamin solution (10x)
Gas phase:
  • 100 % N2

120 METHANOSARCINA MEDIUM

Compound Conc. [g/L] Conc. [mM]
Methanol 15.469 482.765
Na-acetate 2.441 29.761
NaCl 2.197 37.599
NaHCO3 1.953 23.25
Casitone 1.953 -
Yeast extract 1.953 -
NH4Cl 0.488 9.128
MgSO4 x 7 H2O 0.488 1.981
K2HPO4 0.342 1.962
L-Cysteine HCl x H2O 0.293 1.668
Na2S x 9 H2O 0.293 1.22
CaCl2 x 2 H2O 0.244 1.661
KH2PO4 0.225 1.651
HCl 0.002 0.067
FeSO4 x 7 H2O 0.002 0.007
FeCl2 x 4 H2O 0.001 0.007
Sodium resazurin 4.9e-4 0.002
CoCl2 x 6 H2O 1.9e-4 0.001
MnCl2 x 4 H2O 9.8e-5 4.9e-4
Pyridoxine hydrochloride 9.8e-5 4.7e-4
ZnCl2 6.8e-5 5.0e-4
Nicotinic acid 4.9e-5 4.0e-4
p-Aminobenzoic acid 4.9e-5 3.6e-4
(DL)-alpha-Lipoic acid 4.9e-5 2.4e-4
Calcium D-(+)-pantothenate 4.9e-5 2.0e-4
Thiamine HCl 4.9e-5 1.4e-4
Riboflavin 4.9e-5 1.3e-4
Na2MoO4 x 2 H2O 3.5e-5 1.5e-4
NiCl2 x 6 H2O 2.3e-5 1.8e-4
Biotin 2.0e-5 8.0e-5
Folic acid 2.0e-5 4.4e-5
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Solutions:
  • 1 ml Trace element solution SL-10
  • 1 ml Wolin's vitamin solution (10x)
  • 2 ml FeSO4 x 7 H2O solution (0.1% w/v)
Gas phase:
  • 100 % N2
  • 20 % CO2

120a METHANOSARCINA BARKERI MEDIUM

Compound Conc. [g/L] Conc. [mM]
Methanol 15.469 482.765
NaCl 2.197 37.599
Yeast extract 1.953 -
Casitone 1.953 -
NaHCO3 0.83 9.881
NH4Cl 0.488 9.128
MgSO4 x 7 H2O 0.488 1.981
K2HPO4 0.342 1.962
L-Cysteine HCl x H2O 0.293 1.668
Na2S x 9 H2O 0.293 1.22
CaCl2 x 2 H2O 0.244 1.661
KH2PO4 0.225 1.651
HCl 0.002 0.067
FeSO4 x 7 H2O 0.002 0.007
FeCl2 x 4 H2O 0.001 0.007
Sodium resazurin 4.9e-4 0.002
CoCl2 x 6 H2O 1.9e-4 0.001
MnCl2 x 4 H2O 9.8e-5 4.9e-4
Pyridoxine hydrochloride 9.8e-5 4.7e-4
ZnCl2 6.8e-5 5.0e-4
Nicotinic acid 4.9e-5 4.0e-4
p-Aminobenzoic acid 4.9e-5 3.6e-4
(DL)-alpha-Lipoic acid 4.9e-5 2.4e-4
Calcium D-(+)-pantothenate 4.9e-5 2.0e-4
Thiamine HCl 4.9e-5 1.4e-4
Riboflavin 4.9e-5 1.3e-4
Na2MoO4 x 2 H2O 3.5e-5 1.5e-4
NiCl2 x 6 H2O 2.3e-5 1.8e-4
Biotin 2.0e-5 8.0e-5
Folic acid 2.0e-5 4.4e-5
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Solutions:
  • 2 ml FeSO4 x 7 H2O solution (0.1% w/v)
  • 1 ml Trace element solution SL-10
  • 1 ml Wolin's vitamin solution (10x)
Gas phase:
  • 100 % N2
  • 20 % CO2
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