Desulfotomaculum Groll Medium (DSMZ Medium 124a)

[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

Desulfotomaculum Groll Medium
(DSMZ Medium 124a)

培养基配方
Composition per liter:
NaHCO3 .........................................................................4.5g
Na2SO4 ........................................................................2.84g
Na-acetate .....................................................................1.4g
Na-butyrate ....................................................................1.4g
NaCl...........................................................................1.17g
Yeast extract...................................................................1.0g
MgCl2 ·6H2O....................................................................0.4g
KCl.............................................................................0.3g
NH4Cl .........................................................................0.27g
KH2PO4 ........................................................................0.2g
CaCl2 ·2H2O...................................................................0.15g
Resazurin .....................................................................0.5mg
Na2S·9H2O solution...........................................................10.0mL
Substrate solution............................................................10.0mL
Selenite solution.............................................................10.0mL
Vitamin solution..............................................................1.0mL
Trace elements solution ......................................................1.0mL
pH 7.0 ± 0.2 at 25°C
Na2S·9H2O Solution:
Composition per 100.0mL:
Na2S·9H2O.....................................................................3.6g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 100.0mL. Sparge with N2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
Trace Elements Solution:
Composition per liter:
FeCl2 ·4H2O..................................................................1.5g
CoCl2 ·6H2O ..............................................................120.0mg
MnCl2 ·4H2O...............................................................100.0mg
ZnCl2 ......................................................................68.0mg
Na2MoO4 ·2H2O .............................................................24.0mg
NiCl2 ·6H2O................................................................24.0mg
H3BO3 ......................................................................62.0mg
CuCl2 ·2H2O................................................................17.0mg
HCl (0.5M)....................................................................1.0L
Preparation of Trace Elements Solution: Add FeCl2 ·4H2O to
1.0L of 0.5M HCl. Mix thoroughly. Add remaining components. Mix
thoroughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Vitamin Solution:
Thiamine-HCl·2H2O.........................................................10.0mg
p-Aminobenzoic acid.........................................................4.0mg
D (+)-Biotin................................................................1.0mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Selenite Solution:
Composition 10.0mL:
Sodium selenite...........................................................3.0μg
Preparation of Selenite Solution:  Add sodium selenite to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Substrate Solution:
Composition per 10.0mL:
Sodium benzoate...........................................................0.6g
Preparation of Substrate Solution: Add sodium benzoate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Preparation of Medium: Prepare and dispense medium under an
oxygen-free 80% N2 + 20% CO2  gas mixture. Add components, except
NaHCO3 , substrate solution, selenite solution, vitamin solution, and
Na2S·9H2O solution, to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Sparge with 80% N2 + 20% CO2 . Heat gently
and bring to boiling. Cool to room temperature. Add NaHCO3 . Contin-
ue sparging with 80% N2 + 20% CO2  until an equilibrium pH of 6.9-
7.1 is reached. Autoclave for 15 min at 15 psi pressure–121°C. Cool to
25°C while sparging with 80% N2 + 20% CO2 . Aseptically and anaer-
obically add 10.0mL of sterile substrate solution, 10.0mL of sterile sel-
enite solution, 1.0mL of sterile vitamin solution, and 10.0mL of sterile
Na2S·9H2O solution. Mix thoroughly. Aseptically and anaerobically
distribute into sterile tubes or flasks. Alternately the medium can be
distributed to tubes under anaerobic conditions and autoclaved in tubes
prior to addition of substrate solution, selenite solution, vitamin solu-
tion, and Na2S·9H2O solution. Appropriate amounts of these solutions
can then be added to each tube to yield the desired concentrations. Ad-
ditions are performed aseptically and anaerobically under an oxygen-
free 80% N2 + 20% CO2  gas mixture.
Use:  For the cultivation of Desulfotomaculum gibsoniae.