Caloramator proteoclasticus Medium (DSMZ Medium 788)

[所属分类:培养基配方] [发布时间:2020-3-26] [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

Caloramator proteoclasticus Medium
(DSMZ Medium 788)

培养基配方
Composition per liter:
NH4Cl ......................................................2.40g
Na2HPO4 ·2H2O.............................................0.522g
KH2PO4 .....................................................0.48g
MgCl2 ·6H2O................................................0.40g
Yeast extract...............................................0.20g
Resazurin ..................................................0.5mg
NaHCO3  solution ..........................................40.0mL
Na2S·9H2O solution........................................10.0mL
Calcium chloride solution..................................10.0mL
Glucose solution ..........................................10.0mL
Seven vitamin solution......................................1.0mL
Trace elements solution SL-10 ..............................1.0mL
Selenite-tungstate solution.................................1.0mL
pH 7.3 ± 0.2 at 25°C
Glucose Solution:
Composition per 10.0mL:
Glucose ....................................................5.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N2 . Filter sterilize.
Calcium Chloride Solution:
Composition per 10.0mL:
CaCl2 ·2H2O ..............................................0.2g
Preparation of Calcium Chloride Solution:  Add CaCl2 ·2H2O
to distilled/deionized water and bring volume to 10.0mL. Mix thor-
oughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C.
Selenite-Tungstate Solution:
Composition per liter:
NaOH.........................................................0.5g
Na2WO4 ·2H2O...............................................4.0mg
Na2SeO3 ·5H2O..............................................3.0mg
Preparation of Selenite-Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. 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% N2 .
Mix thoroughly. Filter sterilize.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ....................................................10.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
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.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
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 80% N2 + 20% CO2 .
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 . Add components, except seven vitamin solution,
NaHCO3  solution, calcium chloride solution, glucose solution, and
Na2S·9H2O solution, to distilled/deionized water and bring volume to
929.0mL. Mix thoroughly. Sparge with N2 + 20% CO2 . Autoclave for
15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically and anaer-
obically add 1.0mL sterile seven vitamin solution, 40.0mL of sterile
NaHCO3  solution, 10.0mL sterile glucose solution, 10.0mL sterile cal-
cium chloride solution, and 10.0mL of sterile Na2S·9H2O solution.
Mix thoroughly. Adjust pH to 7.2–7.4. Aseptically and anaerobically
distribute into sterile tubes or flasks.
Use:  For the cultivation of Caloramator proteoclasticus.