Desulfitobacterium hafniense Medium

[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

Desulfitobacterium hafniense Medium

培养基配方
Composition per 1005.0mL:
NaHCO3 ......................................................................2.6g
NH4Cl .......................................................................1.0g
Yeast extract................................................................1.0g
K2HPO4 ·3H2O................................................................0.4g
MgCl2 ·6H2O.................................................................0.1g
NaCl.........................................................................0.1g
CaCl2 ·2H2O................................................................0.05g
Resazurin ..................................................................0.5mg
Na2S·9H2O solution.........................................................10.0mL
Sodium pyruvate solution....................................................10.0mL
Wolfe’s vitamin solution...................................................10.0mL
Na2S2O3 solution.............................................................5.0mL
Selenite-tungstate solution..................................................1.0mL
Trace elements solution SL-10 with EDTA......................................1.0mL
pH 7.5 ± 0.2 at 25°C
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O...................................................................0.3g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Sodium Pyruvate Solution:
Composition per 10.0mL:
Sodium pyruvate..............................................................2.5g
Preparation of Sodium Pyruvate Solution:  Add sodium pyru-
vate to distilled/deionized water and bring volume to 10.0mL. Mix
thoroughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl............................................................10.0mg
p-Aminobenzoic acid........................................................5.0mg
Lipoic acid ...............................................................5.0mg
Nicotinic acid.............................................................5.0mg
Riboflavin ................................................................5.0mg
Thiamine·HCl .............................................................5.0mg
Calcium  DL -pantothenate..................................................5.0mg
Biotin ....................................................................2.0mg
Folic acid.................................................................2.0mg
Vitamin B 12 ..............................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize. Sparge with 100% N2 .
Na2S2O3 Solution:
Composition per 10.0mL:
Na2S2O3 ·5H2O.............................................................2.5g
Preparation of Na2S2O3 Solution:  Add Na2S2O3 ·5H2O 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.
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.
Trace Elements Solution SL-10 with EDTA:
Composition per liter:
FeCl2 ·4H2O...............................................................1.5g
Disodium EDTA..............................................................0.5g
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 with EDTA:
Add FeCl2 ·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly.
Preparation of Medium:  Prepare and dispense medium under 80%
N2 + 20% CO2 . Add components, except NaHCO3 , Na2S·9H2O solu-
tion, sodium pyruvate solution, vitamin solution, and Na2S2O3 ·5H2O
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 . Add NaHCO3 . Mix thoroughly. Adjust pH to 7.0. Anaerobically
distribute 9.7mL volumes into anaerobic tubes. Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically and anaerobically add 0.1mL of
sterile Na2S·9H2O solution, 0.1mL of sterile sodium pyruvate solution,
0.1mL of sterile vitamin solution, and 0.05mL of sterile Na2S2O3 ·5H2O
solution to each tube. Mix thoroughly.
Use: For the cultivation of Desulfitobacterium hafniense.