Pelobacter venetianus Marine Medium (DSMZ Medium 296)

[所属分类:培养基配方] [发布时间:2021-7-1] [发布人:网站管理员2] [阅读次数:] [返回]

Pelobacter venetianus Marine Medium
(DSMZ Medium 296)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

NaCl............................................................................................20.0g
MgCl 2 ·6H 2 O .................................................................................3.0g
KCl................................................................................................0.5g
NH 4 Cl .........................................................................................0.25g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Resazurin...................................................................................1.0mg
NaHCO 3  solution.....................................................................10.0mL
Polyethylene glycol solution....................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.36g
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. Auto-
clave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room
temperature.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80%
N 2 + 20% CO 2 . Filter sterilize.
Polyethylene Glycol Solution:
Composition per 10.0mL:
Polyethylene glycol (molecular weight 106–20000)....................1.0g
Preparation of Polyethylene Glycol Solution: Add polyethyl-
ene glycol to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O ...........................................................................190.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O .......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solu-
tion, polyethylene glycol solution, Na 2 S·9H 2 O solution, and trace ele-
ments solution SL-10, to distilled/deionized water and bring volume to
969.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20%
CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and an-
aerobically add 10.0mL NaHCO 3  solution, 10.0mL polyethylene gly-
col solution, 10.0mL Na 2 S·9H 2 O solution, and 1.0mL trace elements
solution SL-10. Mix thoroughly. Aseptically and anaerobically distrib-
ute into sterile tubes or bottles. After inoculation, flush and repressurize
the gas head space of culture bottles with sterile 80% N 2 + 20% CO 2
to 1 bar overpressure.
Use:  For the cultivation of Pelobacter venetianus.