Desulfomicrobium WHB Medium

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulfomicrobium WHB Medium

培养基配方
Composition per 1003.0mL:
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
Solution H..................................................................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
Composition per 870.0mL:
NaCl........................................................................21.0g
MgCl2 ·6H2O ................................................................3.1g
Na2SO4 ......................................................................3.0g
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:
Sodium lactate..............................................................4.0g
Preparation of Solution D:  Add sodium lactate to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution E (Vitamin Solution):
Composition per liter:
Pyridoxine·HCl...........................................................10.0mg
Calcium  DL -pantothenate..................................................5.0mg
Lipoic acid ...............................................................5.0mg
Nicotinic acid.............................................................5.0mg
p-Aminobenzoic acid........................................................5.0mg
Riboflavin ................................................................5.0mg
Thiamine·HCl .............................................................5.0mg
Biotin ....................................................................2.0mg
Folic acid.................................................................2.0mg
Vitamin B 12 ..............................................................0.1mg
Preparation of Solution E (Vitamin Solution): Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Filter sterilize. Sparge with 100% N2 .
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 . Autoclave for 15 min at
15 psi pressure–121°C.
Solution H (Seven Vitamins):
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 H (Seven Vitamins):  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize. Sparge with 100% N2 .
Preparation of Medium: Aseptically and anaerobically combine
solution A with solution B, solution C, solution D, solution E, solution
F, solution G, and solution H, in that order. Mix thoroughly. Anaerobi-
cally distribute into sterile tubes or flasks under 80% N2 + 20% CO2 .
Use: For the cultivation of Desulfomicrobium species.