Desulfonema magnum Medium (DSMZ Medium 202)

[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

Desulfonema magnum Medium
(DSMZ Medium 202)

培养基配方
Composition per 957.6mL:
Solution A..............................................................850.0mL
Solution C...............................................................50.0mL
Solution H...............................................................20.0mL
Solution D...............................................................10.0mL
Solution F (Vitamin solution)............................................10.0mL
Solution I...............................................................10.0mL
Solution G (Artificial sediment)..........................................6.6mL
Solution E ...............................................................1.0mL
Solution B (Trace elements solution SL-10)................................1.0mL
pH 6.9 ± 0.2 at 25°C
Solution A:
NaCl......................................................................21.0g
MgCl2 ·6H2O...............................................................5.5g
Na2SO4 .....................................................................0g
CaCl2 ·2H2O..............................................................1.35g
KCl........................................................................0.5g
KH2PO4 ....................................................................0.2g
NH4Cl .....................................................................0.3g
Resazurin ................................................................0.5mg
Na2SeO3 ·5H2O.............................................................3μg
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
Na2 MoO4 ·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 10.0mL:
Na-acetate...............................................................0.6g
Preparation of Solution D: Add Na-acetate·3H2O to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution E:
Composition per 10.0mL:
Na2 -succinate...........................................................1.0g
Preparation of Solution D: Add Na2 -succinate to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution F (Vitamin Solution):
Composition per liter:
Pyridoxine-HCl.........................................................10.0mg
Vitamin B 12 ...........................................................5.1mg
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
Preparation of Solution F (Vitamin Solution): Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Filter sterilize. Sparge with 100% N2 .
Solution G (Artificial Sediment):
Composition per 6.6mL:
AlCl3 ·6H2O, 4.9% (w/v).................................................5.0mL
Na2CO3 , 10.6% (w/v).....................................................1.6mL
Preparation of Solution G (Artificial Sediment): Combine
components. Mix thoroughly. Sparge with 100% N2 . Autoclave for 15
min at 15 psi pressure–121°C.
Solution H:
Composition per 20.0mL:
Rumen fluid, clarified...................................................20.0mL
Preparation of Solution H: Sparge with 100% N2 . Autoclave for
15 min at 15 psi pressure–121°C.
Solution I:
Composition per 10.0mL:
Na2S·9H2O................................................................0.4g
Preparation of Solution I:  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, 1.0mL solution E, 10.0mL solution F, 6.6mL solution G,
20.0mL solution H, and 10.0mL solution I. Distribute anaerobically
under 80% N2  + 20% CO2 into appropriate vessels. The pH should be
6.9. Addition of 10–20mg sodium dithionite per liter from a 5% (w/v)
solution, freshly prepared under N2  and filter sterilized, may stimulate
growth.
Use: For the cultivation of Desulfonema magnum.