Hippea Medium (DSMZ Medium 854)

[所属分类:培养基配方] [发布时间:2021-4-9] [发布人: ] [阅读次数:] [返回]

Hippea Medium

(DSMZ Medium 854)

培养基配方

Composition per 1010.0mL:
NaCl............................................................................................25.0g
Sulfur, powdered.........................................................................10.0g
Na-acetate.....................................................................................5.0g
MOPS [3-(N-morpholino) propane
sulfonic acid]..........................................................................3.0g
Na 2 S·9H 2 O....................................................................................0.5g
NH 4 Cl .........................................................................................0.33g
CaCl 2 ·2H 2 O ................................................................................0.33g
MgCl 2 ·6H 2 O ...............................................................................0.33g
KCl..............................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
Yeast extract..................................................................................0.1g
Resazurin...................................................................................0.5mg
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
pH 6.1 ± 0.2 at 25°C
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.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................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.
Preparation of Medium: Add components, except vitamin solution, sulfur, and Na 2 S·9H 2 O, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.0. Sparge the medium with 80% N 2 + 20% CO 2 gas mixture for 30 min. Add Na 2 S·9H 2 O. Mix thoroughly. Readjust the pH to 6.0–6.2. Dispense medium under 80% N 2 + 20% CO 2 gas mixture into anaerobe tubes or bottles containing 100.0mg sulfur powder per 10mL medium. Autoclave 20 min at 110°C. Prior to use inject 0.1mL sterile vitamin solution per 10.0mL medium.
Use:  For the cultivation of Hippea maritima.