Carboxydobrachium pacificum Medium (DSMZ Medium 902)

[所属分类:培养基配方] [发布时间:2020-4-7] [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

Carboxydobrachium pacificum Medium
(DSMZ Medium 902)

培养基配方
Composition per 1050mL:
NaCl.........................................................20.0g
MgSO4 ·7H2O..................................................3.9g
KCl...........................................................0.7g
CaCl2 ·2H2O .................................................0.4g
NH4Cl ........................................................0.3g
Na2HPO4 .....................................................0.15g
Yeast extract................................................0.05g
Na2SiO3 .....................................................0.03g
Resazurin....................................................0.5mg
Vitamin solution............................................10.0mL
NaHCO3  solution............................................10.0mL
Na2S·9H2O solution.........................................10.0mL
L -Cysteine solution........................................10.0mL
Pyruvate solution...........................................10.0mL
Trace elements solution SL-10................................1.0mL
pH 7.0 ± 0.2 at 25°C
Pyruvate Solution:
Composition per 10.0mL:
Na-pyruvate ..................................................2.5g
Preparation of Pyruvate Solution:  Add pyruvate to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
L -Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O........................................0.45g
Preparation of  L -Cysteine Solution:  Add  L -cysteine·HCl·H2O 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.................................................0.45g
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.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 .................................................0.5g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be prepared
freshly.
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. Sparge with 100% N2 . Autoclave 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.
Preparation of Medium: Prepare and dispense medium under 100%
N2 . Add components, except vitamin solution, NaHCO3 solution, pyru-
vate solution,  L -cysteine-HCl·H2O solution, and Na2S·9H2O solution,
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 7.0. Sparge with 100% N2 for at least 30 min. Distribute
into anaerobe tubes or bottles. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add per liter, 10.0mL vitamin so-
lution, 10.0mL NaHCO3 solution, 10.0mL pyruvate solution, 10.0mL
L -cysteine-HCl·H2O solution, and 10.0mL Na2S·9H2O. Mix thorough-
ly. The final pH should be 7.0.
Use: For the cultivation of Carboxydibrachium pacificum (Carboxy-
dobrachium pacificum).