Desulfobacterium anilini Medium (DSMZ Medium 476)

[所属分类:培养基配方] [发布时间:2020-6-4] [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

Desulfobacterium anilini Medium
(DSMZ Medium 476)

培养基配方
Composition per 1002.0mL:
Solution A...............................................................870.0mL
Solution C...............................................................100.0mL
Solution D................................................................10.0mL
Solution E (Seven vitamin solution).......................................10.0mL
Solution F................................................................10.0mL
Solution B (Trace elements solution SL-10) ................................1.0mL
Solution G.................................................................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
Composition per 870.0mL:
NaCl........................................................................7.0g
Na2SO4 .....................................................................3.0g
MgCl2 ·6H2O................................................................1.3g
KH2PO4 .....................................................................0.2g
NH4Cl ......................................................................0.3g
KCl.........................................................................0.5g
CaCl2 ·2H2O...............................................................0.15g
Resazurin .................................................................1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly.
Solution B (Trace Elements Solution SL-10):
Composition per liter:
FeCl2 ·4H2O................................................................1.5g
CoCl2 ·6H2O ............................................................190.0mg
MnCl2 ·4H2O.............................................................100.0mg
ZnCl2 ....................................................................70.0mg
Na2MoO4 ·2H2O ...........................................................36.0mg
NiCl2 ·6H2O..............................................................24.0mg
H3BO3 .....................................................................6.0mg
CuCl2 ·2H2O ..............................................................2.0mg
HCl (25% solution)........................................................10.0mL
Preparation of Solution B (Trace Elements Solution SL-10):
Add FeCl2 ·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly. Sparge with 100% N2 . Autoclave for 15 min
at 15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
NaHCO3 .....................................................................5.0g
Preparation of Solution C: Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Flush with 80% N2  + 20% CO2 to remove dissolved oxygen.
Solution D:
Composition per 10.0mL:
Phenol ......................................................................0.1g
Preparation of Solution D: Prepare freshly. Add phenol to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Solution E (Seven Vitamin Solution):
Composition per liter:
Pyridoxine hydrochloride ...................................................300.0mg
Thiamine-HCl·2H2O..........................................................200.0mg
Nicotinic acid..............................................................200.0mg
Vitamin B 12 ...............................................................100.0mg
Calcium pantothenate .......................................................100.0mg
p-Aminobenzoic acid..........................................................80.0mg
D (+)-Biotin.................................................................20.0mg
Solution E (Seven Vitamin Solution): Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution F:
Composition per 10.0mL:
Na2S·9H2O....................................................................0.4g
Preparation of Solution F:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution G (Selenite-Tungstate Solution):
Composition per liter:
NaOH.........................................................................0.5g
Na2WO4 ·2H2O...............................................................4.0mg
Na2SeO3 ·5H2O..............................................................3.0mg
Preparation of Solution G (Selenite-Tungstate Solution):
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Sparge with 100% N2 . Filter sterilize.
Preparation of Medium: Gently heat solution A and bring to boil-
ing. Boil solution A for a few minutes. Cool to room temperature. Gas
with 80% N2  + 20% CO2 gas mixture to reach a pH below 6. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se-
quentially add 1.0mL solution B, 100.0mL solution C, 10.0mL solu-
tion D, 10.0mL solution E, 10.0mL solution F, and 1.0mL solution G.
Distribute anaerobically under 80% N2  + 20% CO2 into appropriate
vessels.
Use: For the cultivation of Desulfobacterium anilini.