Clostridium chartatabidum 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 chartatabidum Medium

培养基配方
Composition per 1001.0mL:
Pancreatic digest of casein..................................................2.0g
Glucose......................................................................0.5g
Glycerol ....................................................................0.5g
Maltose .....................................................................0.5g
Starch, soluble..............................................................0.5g
Yeast extract................................................................0.5g
K2HPO4 ......................................................................0.3g
Hemin ......................................................................1.0mg
Resazurin ..................................................................1.0mg
Rumen fluid...............................................................200.0mL
Cellobiose solution........................................................50.0mL
Mineral solution...........................................................38.0mL
Na2CO3  solution...........................................................30.0mL
L -Cysteine·HCl·H2O solution.............................................10.0mL
Na2S·9H2O solution........................................................10.0mL
pH 6.7 ± 0.2 at 25°C
Cellobiose Solution:
Composition per 50.0mL:
Cellulose, MN 300 or D- Cellobiose..........................................0.5g
Preparation of Cellobiose Solution: Add cellulose or cellobiose
to distilled/deionized water and bring volume to 50.0mL. Mix thor-
oughly. Sparge under 100% CO2 gas for 3 min. Filter sterilize.
Mineral Solution:
Composition per liter:
NaCl.......................................................................12.0g
KH2PO4 .....................................................................6.0g
(NH4)2SO4 ..................................................................6.0g
MgSO4 ·7H2O................................................................2.5g
CaCl2 ·2H2O................................................................0.6g
Preparation of Mineral Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Na2CO3 Solution:
Composition per 30.0mL:
Na2CO3 ....................................................................4.0g
Preparation of Na2CO3 Solution:  Add Na2CO3 to distilled/deion-
ized water and bring volume to 30.0mL. Mix thoroughly. Sparge with
100% CO2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.................................................................0.25g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% CO2 . 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% CO2  for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except cellobiose solu-
tion, Na2CO3  solution, Na2S·9H2O solution, and  L -cysteine·HCl·H2O so-
lution, to distilled/deionized water and bring volume to 900.0mL. Mix
thoroughly. Sparge with 100% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Aseptically and anaerobically add 50.0mL of sterile cellobi-
ose solution, 30.0mL of sterile Na2CO3  solution, 10.0mL of sterile
Na2S·9H2O solution, and 10.0mL of sterile  L -cysteine·HCl·H2O solution.
Mix thoroughly. Aseptically and anaerobically distribute into sterile
screw-capped bottles under 100% CO2 .
Use:  For the cultivation and maintenance of Clostridium chartata-
bidum.