Clostridium cellobioparum Medium

[所属分类:培养基配方] [发布时间:2020-5-9] [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 cellobioparum Medium

培养基配方
Composition per 1010.0mL:
NaCl...........................................................................1.0g
K2HPO4 ........................................................................0.5g
KH2PO4 ........................................................................0.5g
(NH4)2SO4 .....................................................................0.5g
CaCl2 ·2H2O...................................................................0.1g
MgSO4 ·7H2O...................................................................0.1g
Resazurin ....................................................................1.0mg
Rumen fluid, clarified......................................................300.0mL
Cellobiose solution .........................................................50.0mL
NaHCO3  solution.............................................................30.0mL
Na2S·9H2O solution..........................................................20.0mL
L -Cysteine·HCl solution ...................................................10.0mL
pH 6.8 ± 0.2 at 25°C
Cellobiose Solution:
Composition per 50.0mL:
D- Cellobiose.................................................................5.0g
Preparation of Cellobiose Solution: Add cellobiose to distilled/
deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
under 100% N2 gas for 3 min. Filter sterilize. Store under N2 gas.
NaHCO3 Solution:
Composition per 30.0mL:
NaHCO3 .......................................................................10.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 30.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na2S·9H2O Solution:
Composition per 20.0mL:
Na2S·9H2O.................................................................0.25g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized 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 Solution:
Composition per 10.0mL:
L -Cysteine·HCl ..........................................................0.25g
Preparation of  L -Cysteine·HCl Solution: Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave under 100% N2  for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except cellobiose solu-
tion, NaHCO3  solution, Na2S·9H2O solution, and  L -cysteine·HCl solu-
tion, to distilled/deionized water and bring volume to 900.0mL. Mix
thoroughly. Sparge with 100% CO2 . Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically and anaerobically add 50.0mL of sterile
cellobiose solution, 30.0mL of sterile NaHCO3  solution, 20.0mL of
sterile Na2S·9H2O solution, and 10.0mL of sterile  L -cysteine·HCl solu-
tion. Mix thoroughly. Aseptically and anaerobically distribute into
sterile screw-capped bottles.
Use: For the cultivation and maintenance of Clostridium cello-
bioparum and Clostridium polysaccharolyticum.