29a
29a . MEDIUM FOR CHLOROBIUM FERROOXIDANS

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Culture medium recipe

Vol.

×

Main sol. 29a #

ml

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Solution A	460	ml	-	-
Solution B	40	ml	-	-
Solution C	50	ml	-	-
Solution D	450	ml	-	-
Solution E	26	ml	-	-
Solution F	10	ml	-	-
Bicarbonate	10	mM	-	10
FeSO₄	10	mM	1.519	10
Acetate	2	mM	0.118	2
1 Mix solution D, C and E. Bubble with CO₂ in an ice bath under sterile conditions.
2 Fill 50 ml in each bottle of solution A. Before using add 4 ml solution B and 0.1 ml solution F. To final medium add 10 mM bicarbonate, 10 mM FeSO₄ and 2mM acetate.
3 Adjust pH to 6.8.
4 Fill in sterile, N₂ gassed screw-cap tubes under N₂ gas.
5 At 25C in light the incubation time is about 4 weeks.

Solution A #

460

Solution occurs in 2 other media (first defined in Medium 29).

Equipment needed: Autoclave

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
CaCl₂ x 2 H₂O	0.25	g	0.543	3.697
Yeast extract	0.25	g	0.543	-
Distilled water	460.00	ml	-	-
1 Fill 10x 46ml in 100 ml screw-cap bottles. Bubble with N₂/CO₂ and autoclave 121°C 15 min.

Solution B #

135

Solution occurs in 2 other media (first defined in Medium 29).

Equipment needed: Autoclave

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Distilled water	135	ml	-	-
1 Prepare in a screw-cap bottle, bubble with N₂ to replace air, close tightly and autoclave.

Solution C #

50

Solution occurs in 2 other media (first defined in Medium 29).

Equipment needed: Filter

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaHCO₃	1.5	g	30	357.113
H₂O	50.0	ml	-	-
1 Bubble with CO₂ and filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.

Solution D #

450

Solution occurs in 2 other media (first defined in Medium 29).

Equipment needed: Autoclave Ice bath

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Resazurin (0,1%)	0.5	ml	0.005	0.022
Distilled water	450.0	ml	-	-
1 Autoclave in a cotton-stoppered Erlenmeyer flask with an outlet tube for medium, connected to a glass outlet at the bottom of the vessel and has, at the other end, a silicon rubber tube with a pinch cock and a bell for aseptic dispensing of the medium into bottles.
2 Cool to room temperature under an atmosphere of N₂/CO₂ in an ice bath.

Solution E #

25

Solution occurs in 2 other media (first defined in Medium 29).

Equipment needed: Filter

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Ammonium chloride	0.35	g	14	261.731
Ammonium acetate	0.25	g	10	129.735
Pyruvic acid sodium salt	0.25	g	10	90.876
Dextrose	0.25	g	10	50.462
MgSO₄ x 7 H₂O	0.50	g	20	81.144
KCl	0.35	g	14	187.793
KH₂PO₄	0.35	g	14	102.877
Trace element solution SL-10 B	1.00	ml	-	-
Distilled water	25.00	ml	-	-
1 Filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.

Solution F #

100

Solution occurs in 2 other media (first defined in Medium 29).

Equipment needed: Filter Ice bath

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Vitamin B₁₂	0.01	g	0.1	0.074
Distilled water	100.00	ml	-	-
1 Filter sterilized

Trace element solution SL-10 B #

1000

Solution occurs in 1 other media (first defined in Medium ).

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Distilled water	1000.0	ml	-	-
HCl (25%)	7.7	ml	1.925	52.798
FeSO₄ x 7 H₂O	1.5	g	1.5	5.395
ZnCl₂	70.0	mg	0.07	0.514
MnCl₂ x 4 H₂O	100.0	mg	0.1	0.505
H₃BO₃	300.0	mg	0.3	4.851
CoCl₂ x 6 H₂O	190.0	mg	0.19	1.463
CuCl₂ x 2 H₂O	2.0	mg	0.002	0.012
NiCl₂ x 6 H₂O	24.0	mg	0.024	0.185
Na₂MoO₄ x 2 H₂O	36.0	mg	0.036	0.149

Metadata

Last modified:	22.02.22
Source:	DSMZ
Medium type:	Complex medium
Final pH:	6.8
Equipment needed:	Autoclave, Filter, Ice bath

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Associated strains

All strains for this medium:

No strains found

Compound	Conc. [g/L]	Conc. [mM]
FeSO₄	1.519	10
NaHCO₃	1.5	17.856
MgSO₄ x 7 H₂O	0.52	2.11
Ammonium chloride	0.364	6.805
KCl	0.364	4.883
KH₂PO₄	0.364	2.675
Ammonium acetate	0.26	3.373
Pyruvic acid sodium salt	0.26	2.363
Dextrose	0.26	1.312
CaCl₂ x 2 H₂O	0.25	1.701
Yeast extract	0.25	-
Acetate	0.118	2
Resazurin	0.002	0.01
HCl	0.002	0.055
FeSO₄ x 7 H₂O	0.002	0.006
Vitamin B₁₂	1.0e-3	7.4e-4
H₃BO₃	3.1e-4	0.005
CoCl₂ x 6 H₂O	2.0e-4	0.002
MnCl₂ x 4 H₂O	1.0e-4	5.3e-4
ZnCl₂	7.3e-5	5.3e-4
Na₂MoO₄ x 2 H₂O	3.7e-5	1.5e-4
NiCl₂ x 6 H₂O	2.5e-5	1.9e-4
CuCl₂ x 2 H₂O	2.1e-6	1.2e-5
Bicarbonate	-	10
Distilled water	-	-
H₂O	-	-
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phyla	1	1 strain
oxygen	anaerobe	1 strain

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