Desulfobacter sp. Medium (DSMZ Medium 195)

[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

Desulfobacter sp. Medium
(DSMZ Medium 195)

培养基配方
Composition per 991.0mL:
Solution A...............................................................870.0mL
Solution C...............................................................100.0mL
Solution D................................................................10.0mL
Solution E (Vitamin solution).............................................10.0mL
Solution F................................................................10.0mL
Solution B (Trace elements solution SL-10) ................................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
NaCl......................................................................21.0g
MgCl2 ·6H2O...............................................................3.1g
Na2SO4 ....................................................................3.0g
KCl........................................................................0.5g
NH4Cl .....................................................................0.3g
KH2PO4 ....................................................................0.2g
CaCl2 ·2H2O..............................................................0.15g
Resazurin ................................................................1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly.
Solution B (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 Solution B (Trace Elements Solution SL-10):
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. Sparge with 100% N2 . Autoclave for 15 min
at 15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
NaHCO3 ...................................................................5.0g
Preparation of Solution C: Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Flush with 80% N2  + 20% CO2 to remove dissolved oxygen.
Solution D:
Composition per 100.0mL:
Resorcinol................................................................1.1g
Preparation of Solution D: Add resorcinol to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . Filter sterilize. Use freshly prepared.
Solution E (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 .............................................................0.10mg
Preparation of Solution E (Vitamin Solution): Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Solution F:
Composition per 10.0mL:
Na2S·9H2O...................................................................0.4g
Preparation of Solution F:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Gently heat solution A and bring to boil-
ing. Boil solution A for a few minutes. Cool to room temperature. Gas
with 80% N2  + 20% CO2 gas mixture to reach a pH below 6. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se-
quentially add 1.0mL solution B, 100.0mL solution C, 10.0mL solu-
tion D, 10.0mL solution E, and 10.0mL solution F. Distribute
anaerobically under 80% N2  + 20% CO2 into appropriate vessels. Dur-
ing growth the culture is fed once with the same amount of resorcinol
solution.
Use: For the cultivation of unclassified bacterium DSM 4661.