Anoxynatronum Medium (DSMZ Medium 1187)

[所属分类:培养基配方] [发布时间:2019-10-21] [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

Anoxynatronum Medium
(DSMZ Medium 1187)
Composition per liter:
KCl ...............................................................................................0.2g
NH 4 Cl ...........................................................................................0.5g
K 2 HPO 4 .........................................................................................0.2g
MgCl 2 ·6H 2 O .................................................................................0.1g
NaHCO 3  solution.....................................................................50.0mL
Na 2 CO 3  solution.......................................................................50.0mL
Glucose solution ......................................................................50.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Yeast extract solution...............................................................10.0mL
Trace elements solution SL-10 ................................................1.0mL
Vitamin solution ...................................................................... 1.0mL
pH 9.0 ± 0.2 at 25°C
Na 2 CO 3 Solution :
Composition per 50.0mL:
Na 2 CO 3 .......................................................................................25.0g
Preparation of Na 2 CO 3 Solution:  Add Na 2 CO 3 to distilled/de-
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room
temperature.
NaHCO 3 Solution :
Composition per 50.0mL:
NaHCO 3 ......................................................................................25.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3  to distilled/de-
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room
temperature.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract................................................................................. 0.2g
Preparation of Yeast Extract Solution:  Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper-
ature.
Glucose Solution:
Composition per 50.0mL:
Glucose.........................................................................................5.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 50.0mL. 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....................................................................................0.7g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis-
tilled/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.
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 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Add components, except vitamin solu-
tion, yeast extract solution, glucose solution, NaHCO 3  solution,
Na 3 CO 3  solution, and Na 2 S·9H 2 O solution, to distilled/deionized water
and bring volume to 830.0mL. Mix thoroughly. Gently heat and bring
to boiling. Cool to room temperature while sparging with 100% N 2 .
Dispense into tubes or bottles. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C under 100% N 2 . Aseptically and anaerobically
add sterile vitamin solution, yeast extract solution, glucose solution,
NaHCO 3  solution, Na 3 CO 3  solution, and Na 2 S·9H 2 O solution. The fi-
nal pH should be 9.0.
Use: For the cultivation and maintenance of Anoxynatronum spp.