Desulfovibrio gabonensis Medium

[所属分类:培养基配方] [发布时间:2020-6-9] [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

Desulfovibrio gabonensis Medium

培养基配方
Composition per 1002.0mL:
NaCl........................................................................50.0g
MgCl2 ·6H2O.................................................................3.3g
Na2SO4 ......................................................................3.0g
MgSO4 ·7H2O.................................................................1.6g
KCl..........................................................................0.3g
NH4Cl .......................................................................0.3g
KH2PO4 ......................................................................0.2g
CaCl2 ·2H2O.................................................................0.1g
Yeast extract................................................................0.1g
Resazurin ..................................................................0.5mg
Sodium lactate solution....................................................10.0mL
NaHCO3  solution ..........................................................10.0mL
Na2S·9H2O solution........................................................10.0mL
Trace elements solution SL-10 with EDTA.....................................1.0mL
Seven vitamin solution......................................................1.0mL
pH 7.0–7.2 at 25°C
Sodium Lactate Solution:
Composition per 10.0mL:
Sodium lactate...............................................................2.5g
Preparation of Sodium Lactate Solution:  Add sodium lactate 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.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 .....................................................................2.5g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O...................................................................0.2g
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.
Trace Elements Solution SL-10 with EDTA:
Composition per liter:
Disodium EDTA ...............................................................3.0g
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
Preparation of Trace Elements Solution SL-10 with EDTA:
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Adjust pH to 6.0.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine·HCl..............................................................0.3g
Thiamine·HCl................................................................0.2g
Nicotinic acid...............................................................0.2g
Calcium  DL -pantothenate....................................................0.1g
Vitamin B 12 ................................................................0.1g
p-Aminobenzoic acid........................................................80.0mg
Biotin.....................................................................20.0mg
Preparation of Seven Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize. Sparge with 100% N2 .
Preparation of Medium:  Prepare and dispense medium under 80%
N2 % + 20% CO2 . Add components, except sodium lactate solution,
NaHCO3  solution, Na2S·9H2O solution, trace elements solution SL-10
with EDTA, and seven vitamin solution, to distilled/deionized water
and bring volume to 970.0mL. Mix thoroughly. Gently heat and bring
to boiling. Continue boiling for 3 min. Cool to room temperature while
sparging with 80% N2 + 20% CO2 . Anaerobically distribute 9.7mL
volumes into anaerobic tubes. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically add 0.1mL of sterile sodium lactate solution,
0.1mL of sterile NaHCO3  solution, 0.1mL of sterile Na2S·9H2O solu-
tion, 0.01mL of sterile trace elements solution SL-10 with EDTA, and
0.01mL of sterile seven vitamin solution to each tube. Mix thoroughly.
Use: For the cultivation of Desulfovibrio gabonensis.