Desulforhabdus amnigenus Medium (DSMZ Medium 708)

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulforhabdus amnigenus Medium
(DSMZ Medium 708)

培养基配方
Composition per liter:
Na2SO4 .......................................................................2.8g
Na2HPO4 ·2H2O...............................................................0.53g
KH2PO4 ......................................................................0.41g
NH4Cl ........................................................................0.3g
NaCl..........................................................................0.3g
CaCl2 ·2H2O.................................................................0.11g
MgCl2 ·6H2O..................................................................0.1g
Resazurin ...................................................................0.5mg
Na2S·9H2O solution.........................................................10.0mL
Vitamin solution............................................................10.0mL
NaHCO3 solution.............................................................10.0mL
Na-propionate solution .....................................................10.0mL
Selenite-tungstate solution..................................................1.0mL
Seven vitamin solution ......................................................1.0mL
Trace elements solution SL-10................................................1.0mL
pH 7.2–7.6 at 25°C
Na-propionate Solution:
Composition per 10.0mL:
Na-propionate..................................................................2.0g
Preparation of Na-propionate Solution: Add Na-propionate 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.
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 10.0mL:
Na2S·9H2O.......................................................................0.5g
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.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 ........................................................................4.0g
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 . Filter sterilize.
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 . 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.
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.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3  solu-
tion, Na2S·9H2O solution, Na-propionate solution, vitamin solution,
seven vitamin solution, selenite-tungstate solution, and trace elements
solution SL-10, to distilled/deionized water and bring volume to
947.0mL. Mix thoroughly. Adjust pH to 7.2–7.6. Sparge with 80% N2 +
20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anaerobically add 10.0mL NaHCO3  solution, 10.0mL Na2S·9H2O
solution, 10.0mL Na-propionate solution, 10.0mL vitamin solution,
1.0mL seven vitamin solution, 1.0mL selenite-tungstate solution, and
1.0mL trace elements solution SL-10. Mix thoroughly. Aseptically and
anaerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Desulforhabdus amnigena (Desulforhabdus
amnigenus).