[所屬分類:培養(yǎng)基配方] [發(fā)布時(shí)間:2021-4-12] [發(fā)布人: ] [閱讀次數(shù):] [返回]

HNW Medium

(DSMZ Medium 997)

培養(yǎng)基配方

Composition per liter:
DMJ synthetic seawater................................................................1.0L
Vitamin solution.......................................................................10.0mL
NaHCO 3 solution......................................................................10.0mL
NaNO 3 solution.........................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Tungstate solution....................................................................10.0mL
pH 7.2 ± 0.2 at 25°C
Tungstate Solution:
Composition per 10.0mL:
Na 2 WO 4 ·2H 2 O...........................................................................0.1mg
Preparation of Tungstate Solution:  Add Na 2 WO 4 ·2H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
NaHCO 3 Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 20% CO 2 + 80% H 2 . Filter sterilize.
NaNO 3 Solution :
Composition per 10.0mL:
NaNO 3 ...........................................................................................1.0g
Preparation of NaNO 3 Solution:  Add NaNO 3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
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.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
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. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room temperature. Adjust pH to 7.5.
DMJ Synthetic Seawater:
Composition per liter:
NaCl ...........................................................................................30.0g
MgCl 2 ·6H 2 O................................................................................4.18g
MgSO 4 · 7H 2 O................................................................................3.4g
KCl..............................................................................................0.33g
NH 4 Cl ........................................................................................0.25g
K 2 HPO 4 ......................................................................................0.14g
CaCl 2 ·2H 2 O ................................................................................0.14g
Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O............................................................... 0.01g
NiCl 2 ·6H 2 O................................................................................0.5mg
Na 2 SeO 3 ·5H 2 O...........................................................................0.5mg
Trace elements solution SL-10................................................10.0mL
Trace Elements Solution SL-10:
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 SL-10:  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. Adjust pH to 7.0.
Preparation of DMJ Synthetic Seawater:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C . Cool to room temperature.
Preparation of Medium: Aseptically add 10.0mL each of vitamin solution, NaHCO 3 solution, NaNO 3 solution, Na 2 S·9H 2 O solution, and tungstate solution to 1.0L sterile DMJ synthetic seawater. Mix thoroughly. Distribute into tubes. Tightly seal the tubes with butyl rubber stoppers under a gas phase of 80% H 2 + 20% CO 2 (300 kPa).
Use: For the cultivation of Persephonella hydrogeniphila and Hydrogenivirga caldilitoris.