Clostridium thermosuccinogenes Medium

[所属分类:培养基配方] [发布时间:2020-5-18] [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

Clostridium thermosuccinogenes Medium

培养基配方
Composition per 1011.0mL:
Inulin......................................................................5.0g
NaCl........................................................................1.2g
MgCl2 ·6H2O ...............................................................0.4g
KCl.........................................................................0.3g
NH4Cl .....................................................................0.27g
KH2PO4 ....................................................................0.21g
CaCl2 ·2H2O ..............................................................0.15g
Na2SO4 ....................................................................0.1g
Resazurin.................................................................1.0mg
Na2HPO4  solution.........................................................20.0mL
Vitamin solution..........................................................10.0mL
Yeast extract solution....................................................10.0mL
Casamino acids solution...................................................10.0mL
NaHCO3  solution..........................................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
Trace elements solution SL-10..............................................1.0mL
pH 7.0 ± 0.2 at 25°C
Na2HPO4 Solution:
Composition per 20.0mL:
Na2HPO4 ...................................................................2.66g
Preparation of Na2HPO4 Solution:  Add Na2HPO4 to distilled/
deionized water and bring volume to 20.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
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 Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Adjust pH to 7.0. Mix thor-
oughly. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract.............................................................0.03g
Preparation of Yeast Extract Solution:  Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2  for 15 min at 15 psi pressure–121°C.
Casamino Acids Solution:
Composition per 10.0mL:
Casamino acids...........................................................0.03g
Preparation of Casamino Acids Solution:  Add casamino acids
to distilled/deionized water and bring volume to 10.0mL. Mix thor-
oughly. Autoclave under 100% N2  for 15 min at 15 psi pressure–121°C.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 .................................................................1.0mg
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O................................................................0.15mg
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
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 Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Add components, except Na2HPO4  solu-
tion, vitamin solution, yeast extract solution, casamino acids solution,
NaHCO3  solution, Na2S·9H2O solution, and trace elements solution
SL-10, to distilled/deionized water and bring volume to 930.0mL. Mix
thoroughly. Sparge with 80% N2  + 100% CO2 . Autoclave for 15 min at
15 psi pressure–121°C. Aseptically and anaerobically add 20.0mL of
sterile Na2HPO4  solution, 10.0mL of sterile vitamin solution, 10.0mL
of sterile yeast extract solution, 10.0mL of sterile casamino acids solu-
tion, 10.0mL of sterile NaHCO3  solution, 10.0mL of sterile Na2S·9H2O
solution, and 1.0mL of sterile trace elements solution SL-10. Asepti-
cally and anaerobically distribute into tubes or bottles.
Use:  For the cultivation and maintenance of Clostridium thermosucci-
nogenes.