Pseudobutyrivibrio Medium

[所属分类:培养基配方] [发布时间:2021-7-14] [发布人:网站管理员2] [阅读次数:] [返回]

Pseudobutyrivibrio Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

Disodium succinate.......................................................................5.0g
Yeast extract..................................................................................5.0g
NaCl............................................................................................0.45g
(NH 4 ) 2 SO 4 ...................................................................................0.45g
K 2 HPO 4 .....................................................................................0.225g
KH 2 PO 4 .....................................................................................0.225g
MgSO 4 ·7H 2 O..............................................................................0.09g
CaCl 2 ·2H 2 O.................................................................................0.06g
Indigocarmine............................................................................5.0mg
Rumen fluid, clarified............................................................400.0mL
Glucose solution.......................................................................20.0mL
NaHCO 3  solution .....................................................................10.0mL
L -Cysteine·HCl·H 2 O solution ..................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.6–6.8 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% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................6.4g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
L -Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O.....................................................................0.3g
Preparation of  L -Cysteine·HCl·H 2 O Solution:  Add  L -cyste-
ine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi
pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
Preparation of Medium: Add components, except glucose solu-
tion,  L -cysteine·HCl·H 2 O solution, Na 2 S·9H 2 O solution, and NaHCO 3
solution, to distilled/deionized water and bring volume to 960.0mL.
Mix thoroughly. Gently heat and bring to boiling. Cool to room tem-
perature while sparging with 100% CO 2 . Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically and anaerobically add 10.0mL of sterile
glucose solution, 10.0mL of sterile  L -cysteine·HCl solution, 10.0mL of
sterile Na 2 S·9H 2 O solution, and 10.0mL of sterile NaHCO 3  solution.
Mix thoroughly.
Use: For the cultivation of Pseudobutyrivibrio ruminis.