40
40 . PFENNIG'S MEDIUM II WITH SALT

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Culture medium recipe

Vol.

×

Main sol. 40 #

ml

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Medium 29	1000	ml	-	-
NaCl	1	%	10	171.116

Medium 29 #

1000

Solution occurs in 1 other media (first defined in Medium 29).

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	-	-
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.
3 Adjust the pH with filter-sterilised 1M Na₂CO₃ to 6.8 -7.1.
4 Fill in sterile, N₂ gassed screw-cap tubes under N₂ gas.
5 During the first 24 h, the iron of the medium precipitates in the form of black flocks. No other sediment should arise in the otherwise clear medium. Feed periodically with neutralized 3% solution of sodium sulfide to replenish sulfide and with other supplement solutions (see Ref. 3365).

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

Metadata

Last modified:	15.02.22
Source:	DSMZ
Taxonomic range:	Bacteria
Medium type:	Complex medium
Final pH:	n.d.
Equipment needed:	Autoclave, Filter, Ice bath

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Associated strains

All strains for this medium:

9

Compound	Conc. [g/L]	Conc. [mM]
NaCl	10	171.116
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	-
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
Distilled water	-	-
H₂O	-	-
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phyla	2	9 strains
nutrition	phototroph	2 strains
oxygen	anaerobe	8 strains
oxygen	obligate anaerobe	2 strains
temperature	mesophilic	7 strains

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