Desulfobacca Medium (DSMZ Medium 728)

[所属分类:培养基配方] [发布时间:2020-6-4] [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

Desulfobacca Medium
(DSMZ Medium 728)

培养基配方
Composition per liter:
NaHCO3 .......................................................................4.0g
Na2SO4 .......................................................................3.0g
Na-acetate...................................................................1.64g
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
CaCl2  solution..............................................................10.0mL
Trace elements solution SL-10.................................................1.0mL
Selenite-tungstate solution...................................................1.0mL
Seven vitamin solution .......................................................0.2mL
pH 7.1 ± 0.2 at 25°C
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.
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.
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.
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.
CaC2 Solution:
Composition per 10.0mL:
CaCl2 ·2H2O................................................................0.11g
Preparation of CaCl2 Solution:  Add CaCl2 ·2H2O to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80%
N2 + 20% CO2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except Na2S·9H2O
solution, CaCl2  solution, seven vitamin solution, selenite-tungstate so-
lution, and trace elements solution SL-10, to distilled/deionized water
and bring volume to 977.8mL. Mix thoroughly. Adjust pH to 7.0–7.2.
Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Aseptically and anaerobically add 10.0mL Na2S·9H2O so-
lution, 10.0mL CaCl2  solution, 0.2mL 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 Desulfobacca acetoxidans.