​Creatinine/NMH Medium

[所属分类:培养基配方] [发布时间:2020-5-27] [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

Creatinine/NMH Medium

培养基配方
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 B 12 ..............................................................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 components to dis-
tilled/deionized water and bring volume 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 creatinine 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 B 12 ................................................................0.1mg
Preparation of Vitamin Solution : Add components 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 thorough-
ly. 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 dis-
tilled/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, except creatinine solu-
tion, phosphate solution,  L -cysteine·HCl solution, and Na2S·9H2O so-
lution, to distilled/deionized water and bring volume to 880.0mL. Mix
thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 100.0mL of sterile phos-
phate solution, 10.0mL of sterile  L -cysteine·HCl solution, and 10.0mL
of sterile Na2S·9H2O solution. Immediately prior to use, aseptically
and anaerobically add 100.0mL of sterile creatinine solution. Mix thor-
oughly. Aseptically and anaerobically distribute into tubes or bottles.
Use:  For the cultivation and maintenance of Clostridium species.