Clostridium M1 Medium

[所属分类:培养基配方] [发布时间:2020-5-13] [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 M1 Medium

培养基配方
Composition per liter:
NaCl....................................................................10.0g
Betaine·H2O.............................................................6.0g
NaHCO3 ..................................................................5.0g
L -Alanine...............................................................2.2g
NH4Cl ...................................................................1.0g
Yeast extract............................................................1.0g
MgSO4 ·7H2O.............................................................0.5g
CaCl2 ·2H2O...........................................................25.0mg
Resazurin ..............................................................1.0mg
Na2SeO3 ·5H2O........................................................15.0μg
Phosphate solution....................................................100.0mL
Vitamin solution.......................................................10.0mL
Na2S·9H2O solution....................................................10.0mL
Trace elements solution SL-10 ..........................................1.0mL
pH 7.3 ± 0.2 at 25°C
Phosphate Solution:
Composition per 100.0mL:
K2HPO4 ................................................................0.358g
KH2PO4 ................................................................0.223g
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.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O..............................................................0.3g
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.
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. Filter ster-
ilize. Sparge with 100% N2 .
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O...........................................................1.5g
CoCl2 ·6H2O ........................................................190.0mg
MnCl2 ·4H2O.........................................................100.0mg
ZnCl2 ................................................................70.0mg
Na2MoO4 ·2H2O .......................................................36.0mg
NiCl2 ·6H2O..........................................................24.0mg
H3BO3 .................................................................6.0mg
CuCl2 ·2H2O ..........................................................2.0mg
HCl (25% solution)....................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly.
Preparation of Medium: Add components, except phosphate solu-
tion and Na2S·9H2O solution, to distilled/deionized water and bring
volume to 890.0mL. Mix thoroughly. Sparge with 80% N2 + 20% CO2 .
Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaer-
obically add 100.0mL of sterile phosphate solution and 10.0mL of ster-
ile Na2S·9H2O solution. Mix thoroughly. Aseptically and anaerobically
distribute into sterile screw-capped bottles under 80% N2 + 20% CO2 .
Use:  For the cultivation and maintenance of Clostridium halophilum
and Clostridium litorale.