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120d. METHANOSARCINA MARINE MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.35 g 0.342 1.962
2 0.23 g 0.225 1.651
3 0.50 g 0.488 9.128
4 10.00 g 9.766 48.035
5 0.15 g 0.146 0.996
6 23.00 g 22.461 384.325
7
(0.1% w/v in 0.1 N H2SO4)
2.00 ml 0.002 0.007
8 Trace element solution SL-10 1.00 ml - -
9
(0.1% w/v)
0.50 ml 4.9e-4 0.002
10 1.00 g 0.977 9.214
11 Vitamin solution 10.00 ml - -
12 10.00 ml 7.715 240.774
13 0.30 g 0.293 1.668
14 0.30 g 0.293 1.22
15 1000.00 ml 974.812 54110.2
Instructions:
  1. Dissolve ingredients (except carbonate, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Methanol (50% v/v stock solution) and the reducing agents are each autoclaved separately under 100% N2 gas atmosphere as concentrated solutions in tightly closed tubes. Carbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved separately. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Appropriate volumes of the stock solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 6.8 - 7.0, if necessary.
Solution occurs in 366 other media (first defined in medium 141 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 2.0 mg 0.002 0.008
2 2.0 mg 0.002 0.005
3 10.0 mg 0.01 0.049
4 5.0 mg 0.005 0.015
5 5.0 mg 0.005 0.013
6 5.0 mg 0.005 0.041
7 5.0 mg 0.005 0.01
8 0.1 mg 1.0e-4 6.3e-5
9 5.0 mg 0.005 0.036
10 5.0 mg 0.005 0.024
11 1000.0 ml 998.207 55408.8
Solution occurs in 239 other media (first defined in medium 320 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2
(25%; 7.7 M)
10.0 ml 3.17 86.943
3 1.5 g 1.5 7.545
4 70.0 mg 0.07 0.514
5 100.0 mg 0.1 0.505
6 6.0 mg 0.006 0.097
7 190.0 mg 0.19 0.799
8 2.0 mg 0.002 0.012
9 24.0 mg 0.024 0.101
10 36.0 mg 0.036 0.149
11 990.0 ml 988.225 54854.8
Instructions:
  1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.0 ml.

Metadata

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 7.0
Equipment needed: Autoclave, Filter, Hungate tubes

Associated strains

All strains for this medium: 1
Strains with modifications: 1

Gas composition

  • 100 % N2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 985.525 54704.9
NaCl 22.461 384.325
MgCl2 x 6 H2O 9.766 48.035
Methanol 7.715 240.774
Na2CO3 0.977 9.214
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