Clostridium vincentii Medium (DSMZ Medium 769)

[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 vincentii Medium
(DSMZ Medium 769)

培养基配方
Composition per liter:
Yeast extract......................................................................1.0g
Trypticase™........................................................................0.4g
NH4NO3 ............................................................................0.1g
Resazurin ........................................................................0.5mg
Sea water, natural..............................................................300.0mL
NaHCO3  solution ................................................................20.0mL
Phosphate solution...............................................................20.0mL
Lactose solution.................................................................20.0mL
Vitamin solution.................................................................10.0mL
Na2S·9H2O solution..............................................................10.0mL
Cysteine solution................................................................10.0mL
pH 6.5 ± 0.2 at 25°C
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl....................................................................10.0mg
Thiamine-HCl·2H2O.................................................................5.0mg
Riboflavin ........................................................................5.0mg
Nicotinic acid.....................................................................5.0mg
D -Ca-pantothenate.................................................................5.0mg
p-Aminobenzoic acid................................................................5.0mg
Lipoic acid .......................................................................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. Mix thoroughly. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O ..............................................................0.3g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H2O 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. Cool to room temperature.
Lactose Solution:
Composition per 100.0mL:
Lactose............................................................................10.0g
Preparation of Lactose Solution:  Add lactose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Phosphate Solution:
Composition liter:
Na2HPO4 ·12H2O......................................................................43.0g
NaH2PO4 .............................................................................5.44g
Preparation of Phosphate Solution:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Adjust pH to 6.5. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to room temperature.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O.........................................................................0.3g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O 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. Cool
to room temperature.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 .............................................................................5.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3  solu-
tion, Na2S·9H2O solution, lactose solution, vitamin solution, phos-
phate solution, and cysteine solution, to distilled/deionized water and
bring volume to 910.0mL. Mix thoroughly. Adjust pH to 6.8. Sparge
with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 20.0mL NaHCO3  solution,
10.0mL Na2S·9H2O solution, 20.0mL phosphate solution, 10.0mL vi-
tamin solution, 10.0mL cysteine solution, and 20.0mL lactose solution.
Mix thoroughly. Adjust pH to 6.5. Sparge with 80% N2 + 20% CO2 .
Aseptically and anaerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Clostridium vincentii.