Chrysiogenes Medium (DSMZ Medium 818)

[所属分类:培养基配方] [发布时间:2020-4-30] [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

Chrysiogenes Medium
(DSMZ Medium 818)

培养基配方
Composition per liter:
NaCl.....................................................................1.2g
NaHCO3 ..................................................................0.6g
MgCl2 ·6H2O.............................................................0.4g
KCl......................................................................0.3g
NH4Cl ...................................................................0.3g
Na2SO4 ..................................................................0.3g
KH2PO4 ..................................................................0.2g
CaCl2 ·2H2O............................................................0.15g
Resazurin ..............................................................0.5mg
Vitamin solution.......................................................10.0mL
Na-acetate solution ...................................................10.0mL
KNO3 solution .........................................................10.0mL
Seven vitamin solution..................................................1.0mL
Trace elements solution SL-12 ..........................................1.0mL
pH 7.4–7.8 at 25°C
KNO3 Solution:
Composition per 10.0mL:
KNO3 ...................................................................0.5g
Preparation of KNO3 Solution: Add KNO3 to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 . Filter sterilize.
Na-Acetate Solution:
Composition per 10.0mL:
Na-acetate ............................................................0.5g
Preparation of Na-Acetate Solution: Add Na-acetate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Filter sterilize.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ...........................................300.0mg
Thiamine-HCl·2H2O..................................................200.0mg
Nicotinic acid......................................................200.0mg
Vitamin B 12 .......................................................100.0mg
Calcium pantothenate ...............................................100.0mg
p-Aminobenzoic acid..................................................80.0mg
D (+)-Biotin.........................................................20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 .
Mix thoroughly. Filter sterilize.
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.
Trace Elements Solution SL-12:
Composition per liter:
Na2 -EDTA.............................................................5.2g
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-12: 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 thoroughly.
Adjust pH to 6.5. Sparge with 80% N2 + 20% CO2 .
Preparation of Medium: Prepare and dispense medium under 100%
N2 . Add components, except vitamin solution, seven vitamin solution,
KNO3  solution, and Na-acetate solution, to distilled/deionized water
and bring volume to 969.0mL. Mix thoroughly. Sparge with 100% N2 .
Adjust pH to 7.6. Autoclave for 15 min at 15 psi pressure–121°C. Asep-
tically and anaerobically add 10.0mL sterile KNO3  solution, 10.0mL
sterile vitamin solution, 1.0mL sterile seven vitamin solution, and
10.0mL sterile Na-acetate solution. Mix thoroughly. Aseptically and an-
aerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Chrysiogenes arsenatis.