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
Glycine solution.......................................................................50.0mL
Yeast extract solution...............................................................20.0mL
Polysulfide solution .................................................................12.0mL
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
KI.............................................................................................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 tem-
perature. 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.
Yeast Extract Solution:
Composition per 100.0mL:
Yeast extract................................................................................10.0g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Cool to room temperature.
Glycine Solution:
Composition per 100.0mL:
Glycine........................................................................................15.0g
Preparation of Glycine Solution:  Add glycine to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. 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
50.0mL sterile glycine solution, 20.0mL sterile yeast extract solution,
and 12.0mL sterile polysulfide solution to 1.0L sterile base solution.
Mix thoroughly. Aseptically and anaerobically distribute to sterile
tubes or bottles. Do not adjust pH.
Use: For the cultivation of Thermococcus alcaliphilus DSM 10322.