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培养基配方
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Creatinine/NMH Medium
[所属分类:培养基配方] [发布时间:2021-11-15] [发布人:网站管理员2] [阅读次数:] [返回]
Creatinine/NMH Medium
山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com
Composition per 1100.0mL:
Yeast extract........................................................5.0g
NaCl....................................................................1.0g
NaHCO3..............................................................1.0g
MgSO4·7H2O......................................................0.5g
Na2S·9H2O..........................................................0.5g
MnCl2·4H2O .....................................................0.06g
CaSO4·2H2O .....................................................0.05g
FeSO4·7H2O......................................................0.01g
Na2SeO3·5H2O................................................26.0μg
Vitamin B12 .....................................................20.0μg
Resazurin .........................................................1.0mg
Phosphate solution...................................... 100.0mL
Creatinine solution...................................... 100.0mL
Trace elements solution SL-4 ....................... 10.0mL
Vitamin solution............................................ 10.0mL
L-Cysteine·HCl solution ............................... 10.0mL
Na2S·9H2O solution...................................... 10.0mL
pH 7.0 ± 0.2 at 25°C
Phosphate Solution:
Composition per 100.0mL:
K2HPO4.............................................................5.33g
KH2PO4.............................................................2.64g
Preparation of Phosphate Solution: Add com
ponents to distilled/deionized water and bring vol
ume to 100.0mL. Mix thoroughly. Sparge with 100%
N2. Autoclave for 15 min at 15 psi pressure–121°C.
Creatinine Solution:
Composition per 100.0mL:
Creatinine............................................................5.5g
Preparation of Creatinine Solution: Add crea
tinine to distilled/deionized water and bring volume
to 100.0mL. Mix thoroughly. Sparge with 100% N2.
Filter sterilize.
Trace Elements Solution SL-4:
Composition per liter:
EDTA ..................................................................0.5g
FeSO4·7H2O....................................................... 0.2g
Trace elements solution SL-6 ..................... 100.0mL
Trace Elements Solution SL-6:
Composition per liter:
MnCl2·4H2O .......................................................0.5g
H3BO3 .................................................................0.3g
CoCl2·6H2O ........................................................0.2g
ZnSO4·7H2O ...................................................... 0.1g
Na2MoO4·2H2O ............................................... 0.03g
NiCl2·6H2O .......................................................0.02g
CuCl2·2H2O ......................................................0.01g
Preparation of Trace Elements Solution SL-6:
Add components to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly.
Preparation of Trace Elements Solution SL-4:
Add components to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl ..............................................10.0mg
Calcium DL-pantothenate.................................5.0mg
Lipoic acid .......................................................5.0mg
Nicotinic acid...................................................5.0mg
p-Aminobenzoic acid.......................................5.0mg
Riboflavin ........................................................5.0mg
Thiamine·HCl ..................................................5.0mg
Biotin ...............................................................2.0mg
Folic acid..........................................................2.0mg
Vitamin B12 ......................................................0.1mg
Preparation of Vitamin Solution: Add compo
nents to distilled/deionized water and bring volume
to 1.0L. Mix thoroughly.
L-Cysteine·HCl Solution:
Composition per 10.0mL:
L-Cysteine·HCl ...................................................0.5g
Preparation of L-Cysteine·HCl Solution: Add
L-cysteine·HCl to distilled/deionized water and bring
volume to 10.0mL. Mix thoroughly. Sparge with
100% N2. Autoclave for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O..........................................................0.5g
Preparation of Na2S·9H2O Solution: Add
Na2S·9H2O to distilled/deionized water and bring
volume to 10.0mL. Mix thoroughly. Sparge with
100% N2. Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Add components, ex
cept creatinine solution, phosphate solution, L-cys
teine·HCl solution, and Na2S·9H2O solution, to
distilled/deionized water and bring volume to
880.0mL Mix thoroughly. Adjust pH to 7.0. Auto
clave for 15 min at 15 psi pressure–121°C. Aseptical
ly and anaerobically add 100.0mL of sterile
phosphate solution, 10.0mL of sterile L-cysteine·HCl
solution, and 10.0mL of sterile Na2S·9H2O solution.
Immediately prior to use, aseptically and anaerobi
cally add 100.0mL of sterile creatinine solution. Mix
thoroughly. Aseptically and anaerobically distribute
into tubes or bottles.
Use: For the cultivation and maintenance of Clostrid
ium species.