Cellulolytic Medium with Rumen Fluid

[所属分类:培养基配方] [发布时间:2020-4-14] [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

Cellulolytic Medium with Rumen Fluid

培养基配方
Composition per liter:
Basal medium .............................................975.0mL
Alkaline solution..........................................25.0mL
pH 6.8 ± 0.2 at 25°C
Basal Medium:
Composition per 975.0mL:
Agar.......................................................15.0g
NaHCO3 ....................................................6.37g
Pancreatic digest of casein.................................5.0g
Cellobiose..................................................5.0g
NaCl........................................................0.9g
(NH4)2SO4 ..................................................0.9g
K2HPO4 ....................................................0.45g
KH2PO4 ....................................................0.45g
MgSO4 ·7H2O...............................................0.18g
CaCl2 .....................................................0.09g
Resazurin .................................................1.0mg
Rumen fluid, clarified...................................400.0mL
Preparation of Basal Medium: Add components to distilled/de-
ionized water and bring volume to 975.0mL. Mix thoroughly. Gently
heat and bring to boiling under a gas phase of 98% CO2 + 2% H2 . Cool
slightly.
Alkaline Solution:
Composition per 25.0mL:
L -Cysteine·HCl·H2O ........................................0.25g
Na2S·9H2O....................................................0.25g
Preparation of Alkaline Solution: Add components to 25.0mL of
distilled/deionized water. Mix thoroughly. Prepare freshly.
Preparation of Medium: Prepare 975.0mL of basal medium. Heat
to boiling and cool to 25°C. Add 25.0mL of freshly prepared alkaline
solution. Distribute into tubes using anaerobic techniques under a gas
phase of 98% CO2 + 2% H2 . Autoclave for 15 min at 15 psi pressure–
121°C. Adjust pH to 6.8.
Use:  For the cultivation and maintenance of Clostridium polysaccha-
rolyticum.