Desulfotomaculum species Medium II

[所属分类:培养基配方] [发布时间:2020-6-9] [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

Desulfotomaculum species Medium II

培养基配方
Composition per 1001.0mL:
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
pH 7.1–7.4 at 25°C
Solution A:
Composition per 870.0mL:
NaCl........................................................................7.0g
MgCl2 ·6H2O................................................................1.3g
Na2SO4 .....................................................................0.7g
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. 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.
Sparge with 80% N2 + 20% CO2 .
Solution D:
Composition per 10.0mL:
3,4,5-Trimethoxybenzoate...................................................0.42g
Preparation of Solution D:  Add 3,4,5-trimethoxybenzoic acid to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Solution E (Seven Vitamin Solution):
Composition per liter:
Pyridoxine·HCl.............................................................0.3g
Thiamine·HCl ..............................................................0.2g
Nicotinic acid..............................................................0.2g
Calcium  DL -pantothenate...................................................0.1g
Vitamin B 12 ...............................................................0.1g
p-Aminobenzoic acid.......................................................80.0mg
Biotin....................................................................20.0mg
Preparation of Solution E (Seven Vitamin Solution): Add com-
ponents to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pres-
sure–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.
Preparation of Medium: Aseptically and anaerobically combine
solution A with solution B, solution C, solution D, solution E, and so-
lution F, in that order. Mix thoroughly. Anaerobically distribute into
sterile tubes or flasks under 80% N2 + 20% CO2 .
Use: For the cultivation of Desulfotomaculum species.