Desulfobacterium anilini Medium

[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

培养基配方
Composition per 1002.0mL:
Solution A.................................................................870.0mL
Solution C.................................................................100.0mL
Solution D..................................................................10.0mL
Solution E (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
KCl............................................................................0.5g
NH4Cl .........................................................................0.3g
KH2PO4 ........................................................................0.2g
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. Gently heat and
bring to boiling. Continue boiling for 3–4 min. Allow to cool to room
temperature while gassing under 80% N2 + 20% CO2 . Continue gas-
sing until pH reaches below 6.0. Seal the flask under 80% N2 + 20%
CO2 . Autoclave for 15 min at 15 psi pressure–121°C.
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. Gas under 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.
Gas under 80% N2 + 20% CO2 .
Solution D:
Composition per 10.0mL:
Phenol......................................................................0.094g
Preparation of Solution D:  Add phenol to distilled/deionized wa-
ter and bring volume to 10.0mL. Mix thoroughly. Gas under 100% N2 .
Filter sterilize.
Solution E (Vitamin Solution):
Composition per 100.0mL
Nicotinic acid amide........................................................35.0mg
Thiamine dichloride.........................................................30.0mg
p-Aminobenzoic acid.........................................................20.0mg
Biotin......................................................................10.0mg
Calcuim pantothenate .......................................................10.0mg
Pyridoxal·HCl..............................................................10.0mg
Vitamin B 12 ................................................................5.0mg
Preparation of Solution E (Vitamin Solution): Add components
to distilled/deionized water and bring volume to 100.0mL. Mix thor-
oughly. Gas under 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. Gas under 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution G:
Composition per liter:
NaOH.........................................................................0.5g
Na2WO4 ·2H2O...............................................................4.0mg
Na2SeO3 ·5H2O..............................................................3.0mg
Preparation of Solution G:  Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gas under 100% N2 .
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically and anaerobically combine
solution A with solution B, solution C, solution D, solution E, solution
F, and solution G, in that order. Mix thoroughly. Anaerobically distrib-
ute into sterile tubes or flasks under 80% N2 + 20% CO2
Use:  For the cultivation and maintenance of Desulfobacterium anilini.