Desulfococcus Medium

[所属分类:培养基配方] [发布时间:2020-6-5] [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

Desulfococcus Medium

培养基配方
Composition per 1001.0mL:
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
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
Na2SeO3 ·5H2O.............................................................3.0μg
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:
Sodium benzoate...............................................................0.6g
Preparation of Solution D:  Add sodium benzoate to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Gas under
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 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.
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.
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 and maintenance of Desulfococcus multiv-
orans.