Desulfovigra adipica Medium (DSMZ Medium 868)

[所属分类:培养基配方] [发布时间:2020-6-10] [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

Desulfovigra adipica Medium
(DSMZ Medium 868)

培养基配方
Composition per 2.0L:
Solution A................................................................940.0mL
Solution E ................................................................50.0mL
Solution K.................................................................50.0mL
Solution G.................................................................20.0mL
Solution F.................................................................10.0mL
Solution M (Vitamin solution)..............................................10.0mL
Solution H..................................................................7.0mL
Solution B..................................................................1.0mL
Solution C..................................................................1.0mL
Solution D..................................................................1.0mL
Solution I .................................................................1.0mL
Solution J (Trace elements solution SL-10) .................................1.0mL
Solution L (Selenite-tungstate solution)....................................1.0mL
Solution N................................................................variable
pH 7.1 ± 0.2 at 25°C
Solution A:
Composition per 940.0mL:
NaCl.........................................................................1.0g
KCl..........................................................................0.5g
MgCl2 ·6H2O ................................................................0.4g
NH4Cl ......................................................................0.25g
KH2PO4 ......................................................................0.2g
CaCl2 ·2H2O ...............................................................0.15g
Resazurin..................................................................0.5mg
Preparation of Solution A: Prepare under 80% N2 + 20% CO2 gas
atmosphere. Add components to distilled/deionized water and bring vol-
ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N2
+ 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
25°C.
Solution B:
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: Add FeCl2 ·4H2O to 10.0mL of HCl so-
lution. Mix thoroughly. Add distilled/deionized water and bring vol-
ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with
80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 25°C.
Solution C:
Composition per liter:
Pyridoxine hydrochloride...................................................300.0mg
Thiamine-HCl·2H2O.........................................................200.0mg
Nicotinic acid.............................................................200.0mg
Vitamin B 12 ..............................................................100.0mg
Calcium pantothenate ......................................................100.0mg
p-Aminobenzoic acid.........................................................80.0mg
D (+)-Biotin................................................................20.0mg
Preparation of Solution C: Add components to distilled/deionized
water and bring volume to 1.0L. Sparge with 100% N2 . Mix thorough-
ly. Filter sterilize.
Solution D:
Composition per liter:
NaOH..........................................................................0.5g
Na2WO4 ·2H2O................................................................4.0mg
Na2SeO3 ·5H2O...............................................................3.0mg
Preparation of Solution D: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Sparge with 100%
N2 . Filter sterilize.
Solution E:
Composition per 100.0mL:
NaHCO3 .......................................................................5.0g
Preparation of Solution E:  Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 25°C.
Solution F:
Composition per 10.0mL:
Yeast extract..................................................................1.0g
Preparation of Solution F:  Add yeast extract to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 25°C.
Solution G :
Composition per 20.0mL:
Na2S·9H2O...................................................................0.625g
Preparation of  Solution G : Add Na2S·9H2O to distilled/deionized
water and bring volume to 20.0mL. Mix thoroughly. Autoclave under
100% N2 for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Solution H:
Composition per 10.0mL:
Na2SO4 ......................................................................1.0g
Preparation of Solution H:  Add Na2SO4 to distilled/deionized wa-
ter and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N2
gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to
25°C.
Solution I:
Composition per 10.0mL:
Propanol......................................................................1.0g
Preparation of Solution I:  Add propanol to distilled/deionized wa-
ter and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N2
gas mixture. Filter sterilize.
Solution J (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 J (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 80% N2 + 20% CO2 . Autoclave
for 15 min at 15 psi pressure–121°C.
Solution K:
Composition per 100.0mL:
NaHCO3 .......................................................................5.0g
Preparation of Solution K:  Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 25°C.
Solution L (Selenite–Tungstate Solution):
Composition per liter:
NaOH...........................................................................0.5g
Na2WO4 ·2H2O.................................................................4.0mg
Na2SeO3 ·5H2O................................................................3.0mg
Preparation of Solution L (Selenite–Tungstate Solution):
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Sparge with 100% N2 . Filter sterilize.
Solution M (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 ..................................................................0.1mg
Preparation of Solution M (Vitamin Solution):  Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Sparge with 80% H2 + 20% CO2 . Filter sterilize.
Solution N:
Composition per 100.0mL:
Na2CO3 ........................................................................5.0g
Preparation of Solution N:  Add Na2CO3 to distilled/deionized wa-
ter and bring volume to 100.0mL. Mix thoroughly. Sparge with 100%
N2 gas mixture. Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Sequentially add 1.0mL solution B,
1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so-
lution F, 20.0mL solution G, 7.0mL solution H, 1.0mL solution I,
1.0mL solution J, 50.0mL solution K, 1.0mL solution L, and 10.0mL
solution M, to 940.0mL solution A. Mix thoroughly. Adjust pH to 7.1
with solution N. Distribute anaerobically under 80% N2  + 20% CO2
into appropriate vessels.
Use: For the cultivation of Desulfovirga adipica (Desulfobacterium
sp.)