Clostridium caminithermalis Medium (DSMZ Medium 986)

[所属分类:培养基配方] [发布时间:2020-5-8] [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 caminithermalis Medium
(DSMZ Medium 986)

培养基配方
Composition per liter:
Sea salt (Sigma) ..................................................30.0g
Glucose ............................................................4.0g
NH4Cl ..............................................................1.0g
Peptone.............................................................0.5g
Yeast extract.......................................................0.5g
KH2PO4 .............................................................0.3g
K2HPO4 .............................................................0.3g
Resazurin .........................................................1.0mg
Glucose solution ................................................100.0mL
Vitamin solution..................................................10.0mL
L -Cysteine·HCl solution.........................................10.0mL
NaHCO3  solution .................................................10.0mL
Na2S·9H2O solution...............................................10.0mL
pH 7.0 ± 0.2 at 25°C
Glucose Solution :
Composition per 100.0mL:
Glucose ..........................................................4.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . Filter sterilize.
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.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl ...............................................0.25g
Preparation of  L -Cysteine Solution : Add  L -cysteine·HCl to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave under 100% N2 for 15 min at 15 psi
pressure–121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl...................................................10.0mg
Thiamine-HCl·2H2O................................................5.0mg
Riboflavin........................................................5.0mg
Nicotinic acid....................................................5.0mg
D -Ca-pantothenate ...............................................5.0mg
p-Aminobenzoic acid...............................................5.0mg
Lipoic acid.......................................................5.0mg
Biotin............................................................2.0mg
Folic acid .......................................................2.0mg
Vitamin B 12 ....................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
NaHCO 3 Solution :
Composition per 10.0mL:
NaHCO3 .........................................................2.0g
Preparation of NaHCO3 Solution:  Add components to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO2 + 80% H2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature.
Preparation of Medium: Add components, except vitamin, glu-
cose, bicarbonate, cysteine and sulfide solutions, to distilled/deionized
water and bring volume to 860.0mL. Mix thoroughly. Gently heat and
bring to boiling. Boil for 3 min. Cool while sparging with 100% N2 .
Dispense  under  100% N2  gas atmosphere.  Autoclave under 100% N2
for 15 min at 15 psi pressure–121°C. Aseptically and anoxically add
vitamin, glucose, bicarbonate, cysteine, and sulfide solutions. Adjust
final pH of the medium to pH 7.0.
Use:  For the cultivation of Clostridium caminithermalis.