Heterotrophic Hyperthermophilic Archaea Medium

[所属分类:培养基配方] [发布时间:2021-4-6] [发布人: ] [阅读次数:] [返回]

Heterotrophic Hyperthermophilic

Archaea Medium

培养基配方

Composition per liter:
Pancreatic digest of casein............................................................3.0g
Glucose .........................................................................................3.0g
Yeast extract..................................................................................3.0g
Artificial seawater..................................................................990.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 7.2 ± 0.2 at 25°C
Artificial Seawater:
Composition per liter:
NaCl............................................................................................20.0g
MgSO 4 ·7H 2 O................................................................................6.0g
MgCl 2 ·6H 2 O .................................................................................3.0g
(NH 4 ) 2 SO 4 ....................................................................................1.0g
NaHCO 3 ........................................................................................0.2g
CaCl 2 ·2H 2 O ..................................................................................0.3g
KCl................................................................................................0.5g
KH 2 PO 4 .......................................................................................0.42g
NaBr............................................................................................0.05g
SrCl 2 ·6H 2 O .................................................................................0.02g
Fe(NH 4 ) citrate ...........................................................................0.01g
Wolfe’s mineral solution............................................................5.0mL
Vitamin solution.........................................................................5.0mL
Wolfe’s Mineral 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
CoCl 2 ·6H 2 O..................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
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
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add remaining components one at a time. Add distilled/deionized water to 1.0L. Adjust pH to 6.8.
Vitamin Solution:
Composition per liter:
Niacin.......................................................................................10.0mg
Pantothenate.............................................................................10.0mg
Lipoic acid...............................................................................10.0mg
p-Aminobenzoic acid...............................................................10.0mg
Thiamine (B 1 )..........................................................................10.0mg
Riboflavin (B 2 ) ........................................................................10.0mg
Pyridoxine (B 6 )........................................................................10.0mg
Cobalamin (B 12 ).......................................................................10.0mg
Biotin.........................................................................................4.0mg
Folic acid ...................................................................................4.0mg
Preparation of Vitamin Solution:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Artificial Seawater:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.8g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Before use, neutralize to pH 7.0 with sterile HCl.
Preparation of Medium: Add components, except Na 2 S·9H 2 O solution, to artificial seawater and bring volume to 990.0mL. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 for 20 min. Aseptically and anaerobically add 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly. Adjust medium pH to 7.2 by adding sterile anaerobic HCl. Aseptically and anaerobically distribute into sterile tubes or bottles.
Use: For the cultivation of hyperthermophilic archaea.