Desulfonema limicola Medium (DSMZ Medium 201)

[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 limicola Medium
(DSMZ Medium 201)

培养基配方
Composition per 1007.6mL:
Solution A..................................................................850.0mL
Solution C..................................................................100.0mL
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 7.6 ± 0.2 at 25°C
Solution A:
NaCl..........................................................................13.0g
Na2SO4 ........................................................................3.0g
MgCl2 ·6H2O...................................................................2.2g
KCl............................................................................0.5g
KH2PO4 ........................................................................0.2g
NH4Cl .........................................................................0.3g
CaCl2 ·2H2O..................................................................0.15g
Resazurin ....................................................................0.5mg
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 10.0mL:
Na-acetate·3H2O..............................................................2.5g
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 E: 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
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 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 com-
ponents. 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
7.6. 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 limicola.