Caloramator viterbensis Medium (DSMZ Medium 947)

[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 viterbensis Medium
(DSMZ Medium 947)

培养基配方
Composition per liter:
Glycerol .....................................................3.0g
KH2PO4 .......................................................2.0g
NaHCO3 .......................................................1.0g
(NH4)2SO4 ....................................................0.5g
NH4Cl ........................................................0.5g
Yeast extract.................................................0.3g
MgCl2 ·6H2O.................................................0.04g
CaCl2 ·2H2O.................................................0.04g
Resazurin ...................................................0.5mg
Cysteine solution............................................10.0mL
Na2S·9H2O solution..........................................10.0mL
Trace elements solution SL-10 ................................1.0mL
Selenite-tungstate solution...................................1.0mL
Vitamin solution..............................................0.2mL
pH 6.3 ± 0.2 at 25°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.
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.
Na2S·9H2O Solution:
Composition per 100.0mL:
Na2S·9H2O..........................................................1.3g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O 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.
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% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Cysteine Solution:
Composition per 100.0mL:
L -Cysteine·HCl·H2O..............................................1.3g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H2O to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3 ,
Na2S·9H2O solution, and cysteine solution, to distilled/deionized water
and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to
boiling. Cool to room temperature while sparging with 80% N2 + 20%
CO2 . Add solid NaHCO3 . Adjust pH to 6.3 by equilibrating with the
same gas mixture. Distribute to tubes or bottles. Autoclave for 15 min at
15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL ster-
ile cysteine solution and 10.0mL sterile Na2S·9H2O solution. Mix thor-
oughly. Adjust pH to 6.8. Aseptically and anaerobically distribute into
sterile tubes or bottles.
Use:  For the cultivation of Caloramator viterbiensis.