Artificial Seawater Medium (DSMZ Medium 1010)

[所属分类:培养基配方] [发布时间:2019-11-11] [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

Artificial Seawater Medium
(DSMZ Medium 1010)
Composition per liter:
NaCl............................................................................................26.4g
MgSO4 ·7H2O................................................................................6.8g
MgCl2 ·6H2O .................................................................................5.7g
CaCl2 ·2H2O ................................................................................1.47g
KCl..............................................................................................0.66g
Resazurin ......................................................................................0.5g
K2HPO4 .........................................................................................0.2g
KBr .............................................................................................0.09g
Na2S·9H2O solution.................................................................10.0mL
Vitamin solution.......................................................................10.0mL
Sodium lactate solution............................................................10.0mL
Ammonium chloride solution..................................................10.0mL
Bicarbonate solution................................................................10.0mL
Dihydrogen phosphate solution...............................................10.0mL
Seven vitamin solution ..............................................................1.0mL
Selenite/tungstate solution.........................................................1.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.3 ± 0.2 at 25°C
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................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: Add FeCl 2 ·4H 2 O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 100% N 2 . 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.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na2S·9H2O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
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 Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H 2 + 20% CO 2 . Filter sterilize.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................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 Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 .
Mix thoroughly. Filter sterilize.
Dihydrogen Phosphate Solution:
Composition per 10.0mL:
KH2PO4 .........................................................................................0.2g
Preparation of Dihydrogen Phosphate Solution: Add KH 2 PO 4
to distilled/deionized water and bring volume to 10.0L. Sparge with
100% N 2 . Mix thoroughly. Filter sterilize.
Sodium Lactate Solution:
Composition per 10.0mL:
Sodium lactate...............................................................................2.3g
Preparation of Sodium Lactate Solution: Add sodium lactate to
distilled/deionized water and bring volume to 10.0L. Sparge with
100% N 2 . Mix thoroughly. Filter sterilize.
Bicarbonate Solution:
Composition per 10.0mL:
NH4Cl .........................................................................................0.25g
Preparation of Bicarbonate Solution: Add NH 4 Cl to distilled/
deionized water and bring volume to 10.0L. Sparge with 100% N 2 . Mix
thoroughly. Filter sterilize.
Ammonium Chloride Solution:
Composition per 10.0mL:
NaHCO3 ........................................................................................2.5g
Preparation of Ammonium Chloride Solution: Add NaHCO 3
to distilled/deionized water and bring volume to 10.0L. Sparge with
100% N 2 . Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except dihydrogen
phosphate, ammonium chloride, bicarbonate, lactate, vitamin, and sul-
fide solutions, to distilled/deionized water and bring volume to
940.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 3
min. Cool to room temperature while sparging with 20% CO 2 + 80%
N 2 . Dispense into tubes or bottles under atmosphere of 20% CO 2 +
80% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C
under an atmosphere of 20% CO 2 + 80% N 2 . Aseptically and anaero-
bically add sterile dihydrogen phosphate, ammonium chloride, bicar-
bonate, lactate, vitamin, and sulfide solutions. Adjust final pH to 7.3.
Use: For the cultivation and maintenance of Desulfobacterium cor-
rodens