[所屬分類:培養(yǎng)基配方] [發(fā)布時(shí)間:2020-7-15] [發(fā)布人:] [閱讀次數(shù):] [返回]

山東拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Fervidobacterium Medium

培養(yǎng)基配方
Composition per liter:
Pancreatic digest of casein...............................................10.0g
Glucose....................................................................5.0g
Yeast extract..............................................................3.0g
K2HPO4 ....................................................................1.5g
NH4Cl .....................................................................0.9g
KH2PO4 ...................................................................0.75g
MgCl2 ·6H2O ..............................................................0.2g
Na2S·9H2O solution......................................................10.0mL
Trace elements solution..................................................9.0mL
Wolfe’s vitamin solution................................................5.0mL
Resazurin (0.2% solution) ...............................................1.0mL
FeSO4 ·7H2O (10% solution).............................................0.03mL
pH 7.0 ± 0.1 at 25°C
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid..................................................12.5g
NaCl....................................................................1.0g
FeCl3 ·4H2O............................................................0.2g
CaCl2 ·2H2O ...........................................................0.1g
MnCl2 ·4H2O ...........................................................0.1g
ZnCl2 ..................................................................0.1g
CuCl2 .................................................................0.02g
Na2SeO3 ...............................................................0.02g
CoCl2 ·6H2O..........................................................0.017g
H3BO3 .................................................................0.01g
Na2MoO4 ·2H2O.........................................................0.01g
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Filter sterilize. Maintain under an atmosphere of 100%
N2 .
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl.....................................................0.01g
Thiamine·HCl.......................................................5.0mg
Riboflavin..........................................................5.0mg
Nicotinic acid......................................................5.0mg
Calcium pantothenate ...............................................5.0mg
p-Aminobenzoic acid.................................................5.0mg
Thioctic acid.......................................................5.0mg
Biotin..............................................................2.0mg
Folic acid .........................................................2.0mg
Cyanocobalamin......................................................0.1mg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize. Maintain under an atmosphere of 100% N2 .
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O...........................................................0.5g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize. Maintain under an atmosphere of 100% N2 .
Preparation of Medium: Add components, except sodium sulfide
solution, trace elements solution, and Wolfe’s vitamin solution, to dis-
tilled/deionized water and bring volume to 976.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool under an atmo-
sphere of 100% N2 . Aseptically add 9.0mL of trace elements solution
and 5.0mL of Wolfe’s vitamin solution under an atmosphere of 100%
N2 . Mix thoroughly. Aseptically distribute into sterile tubes or flasks
under an atmosphere of 100% N2 . Add Na2S·9H2O solution just prior
to use to a concentration of 0.1%.
Use:  For the cultivation and maintenance of Clostridium species, Fer-
vidobacterium nodosum, Fervidobacterium islandicum, and Thermoa-
naerobium brockii.