Alkaliphilic Thermococcus Medium (DSMZ Medium 926)

[所属分类:培养基配方] [发布时间:2019-9-29] [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

Alkaliphilic Thermococcus Medium
(DSMZ Medium 926)
Composition per 1082.0mL:
Base solution........................................................................1000.0mL
Casamino acids solution..........................................................20.0mL
Polysulfide solution...................................................................8.0mL
Base Solution:
Composition per 2000.0mL:
NaCl............................................................................................27.7g
MgSO 4 ·7H 2 O................................................................................7.0g
MgCl 2 ·6H 2 O .................................................................................5.5g
K 2 HPO 4 .........................................................................................1.0g
KCl..............................................................................................0.65g
NaHCO 3 ......................................................................................0.32g
NaBr..............................................................................................0.1g
H 3 BO 3 .........................................................................................0.03g
KJ.............................................................................................15.0mg
CaCl 2 ·2H 2 O .............................................................................0.05mg
Trace elements solution...........................................................20.0mL
Preparation of Base Solution:  Sparge 2.0L of distilled/deionized
water with 100% N 2  to remove O 2 . Add components to 2000.0mL of O 2 -
free distilled/deionized water. Mix thoroughly. Sparge with 100% N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room tempera-
ture. Do not adjust the pH.
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O ..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution:  Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Polysulfide Solution:
Composition per 100.0mL:
Na 2 S·9H 2 O....................................................................................1.2g
Sulfur ..........................................................................................0.16g
Preparation of Polysulfide Solution: Sparge 100.0mL distilled/
deionized water with 100% N 2 . Add Na 2 S·9H 2 O. Mix thoroughly. Add
sulfur. The solution will be dark yellow. Sparge with 100% N 2 . Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Casamino Acids Solution:
Composition per 100.0mL:
Casamino acids...........................................................................10.0g
Preparation of Casamino Acids Solution:  Add casamino acids
to distilled/deionized water and bring volume to 100.0mL. Mix thor-
oughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature.
Preparation of Medium:  Aseptically and anaerobically add
20.0mL sterile casamino acids solution, and 12.0mL sterile polysulfide
solution to 1.0L sterile base solution. Mix thoroughly. Aseptically and
anaerobically distribute to sterile tubes or bottles. There may be precip-
itation of material and the medium will turn pale yellow due to the ad-
dition of the polysulfide. The color will disappear as the strain grows.
Do not adjust pH.
Use: For the cultivation of Thermococcus acidaminovorans DSM
11096.