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Clostridium grantii Medium

[所属分类:培养基配方] [发布时间:2020-5-11] [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 grantii Medium

培养基配方
Composition per liter:
NaCl.....................................................................19.45g
MgCl2 .....................................................................8.8g
Peptone ...................................................................5.0g
Na2SO3 ...................................................................3.24g
NaHCO3 ....................................................................2.0g
CaCl2 .....................................................................1.8g
Yeast extract..............................................................1.0g
KCl.......................................................................0.55g
Ferric citrate.............................................................0.1g
KBr.......................................................................0.08g
SrCl2 ....................................................................0.03g
H3BO3 ....................................................................0.02g
Na2HPO4 ..................................................................8.0mg
Na2SiO3 ..................................................................4.0mg
NaF.......................................................................2.4mg
NH4NO3 ...................................................................1.6mg
Resazurin ................................................................0.5mg
Glucose solution ........................................................20.0mL
L -Cysteine·HCl·H2O solution ..........................................10.0mL
Na2S·9H2O solution......................................................10.0mL
NaHCO3  solution ........................................................10.0mL
pH 6.6 ± 0.2 at 25°C
Glucose Solution:
Composition per 20.0mL:
D -Glucose................................................................5.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 20.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
L -Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O .....................................................0.3g
Preparation of  L -Cysteine·HCl·H2O Solution:  Add  L -cyste-
ine·HCl·H2O 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.
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.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 ....................................................................0.16g
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.
Preparation of Medium: Add components, except glucose solu-
tion,  L -cysteine·HCl·H2O solution, Na2S·9H2O solution, and NaHCO3
solution, to distilled/deionized water and bring volume to 950.0mL.
Mix thoroughly. Gently heat and bring to boiling. Cool to room tem-
perature while sparging with 80% N2 + 20% CO2 . Anaerobically dis-
tribute 9.5mL volumes into anaerobic tubes. Autoclave for 15 min at
15 psi pressure–121°C. Aseptically add 0.2mL of sterile glucose solu-
tion, 0.1mL of sterile  L -Cysteine·HCl·H2O solution, 0.1mL of sterile
Na2S·9H2O solution, and 0.1mL of sterile NaHCO3  solution to each
tube. Mix thoroughly.
Use: For the cultivation of Clostridium grantii.

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