Chromatium salexigens Medium

[所属分类:培养基配方] [发布时间:2020-4-29] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Chromatium salexigens Medium

培养基配方
Composition per 4990.0mL:
Solution A..............................................................4000.0mL
Solution B...............................................................860.0mL
Solution C (Vitamin B 12 solution) ........................................5.0mL
Solution D (Trace elements solution SL-12B)................................5.0mL
Solution E ..............................................................100.0mL
Solution F................................................................20.0mL
pH 7.3 ± 0.2 at 25°C
Solution A:
Composition per 4000.0mL:
NaCl......................................................................100.0g
MgCl2 ·6H2O................................................................3.0g
MgSO4 ......................................................................2.5g
KH2PO4 .....................................................................1.7g
NH4Cl ......................................................................1.7g
KCl.........................................................................1.7g
CaCl2 ·2H2O...............................................................1.25g
Sodium acetate..............................................................0.5g
Na2S2O3 ....................................................................0.5g
Preparation of Solution A : Add components to distilled/deionized
water and bring volume to 4.0L. Mix thoroughly. Adjust pH to 6.0.
Dispense into a 5.0L flask with four openings at the top (two openings
are in a central silicon rubber stopper and two openings are gas-tight
screw caps). Add a teflon-coated magnetic stir bar to the flask. Auto-
clave for 45 min at 15 psi pressure–121°C. Cool to room temperature
under 100% N2 at 0.05–0.1 atm pressure (use a manometer to measure
low pressure).
Solution B:
Composition per 860.0mL:
Distilled/deionized water...............................................860.0mL
Preparation of Solution B : Add 860.0mL of distilled/deionized
water to a cotton-stoppered flask. Autoclave for 20 min at 15 psi–
121°C. Cool to room temperature under 100% N2 in an anaerobic jar.
Solution C (Vitamin B 12 Solution):
Composition per 5.0mL:
Vitamin B 12 .............................................................1.0mg
Preparation of Solution C (Vitamin B 12 Solution): Add vita-
min B 12 to distilled/deionized water and bring volume to 5.0mL. Mix
thoroughly. Filter sterilize.
Solution D (Trace Elements Solution SL-12B):
Composition per liter:
Disodium ethylendiamine-tetraacetate
(Disodium EDTA)........................................................3.0g
FeSO4 ·7H2O...........................................................1.1g
H3BO3 .................................................................0.3g
CoCl2 ·6H2O..........................................................0.19g
MnCl2 ·2H2O ........................................................50.0mg
ZnCl2 ...............................................................42.0mg
NiCl2 ·6H2O.........................................................24.0mg
Na2MoO4 ·2H2O.......................................................18.0mg
CuCl2 ·2H2O..........................................................2.0mg
Preparation of Solution D (Trace Elements Solution SL-12B):
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi–121°C.
Solution E:
Composition per 100.0mL:
NaHCO3 ................................................................7.5g
Preparation of Solution E : Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% CO2 until saturated. Filter sterilize under 100% CO2 in a sterile,
gas-tight 100.0mL screw-capped bottle.
Solution F:
Composition per 100.0mL:
Na2S·9H2O.............................................................10.0g
Preparation of Solution F : Add Na2S·9H2O to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Dispense into a
screw-capped bottle. Sparge with 100% N2  for 3–4 min. Autoclave for
15 min at 15 psi pressure–121°C.
Preparation of Medium: Saturate cooled solution A under 100%
CO2 at 0.05–0.1 atm pressure for 30 min with magnetic stirring. Add
860.0mL of solution B, 5.0mL of solution C, 5.0mL of solution D,
100.0mL of solution E, and 20.0mL of solution F through one of the
screw-capped openings under 95% N2  and 5%  CO2 with magnetic stir-
ring. Adjust pH to 7.3 with sterile 2M HCl or sterile 2M Na2CO3 solu-
tion. Aseptically and anaerobically distribute the medium through the
medium outlet tube into sterile 100.0mL bottles under 95% N2  and 5%
CO2 at 0.05–0.1 atm pressure. Leave a small gas bubble in each bottle
to accommodate pressure changes. After 24 hr, the iron in the medium
will precipitate out of solution as black flocs.
Use:  For the growth and maintenance of Chromatium salexigens.