Rhodovulum iodosum Rhodovulum robiginosum Medium (DSMZ Medium 929)

[所属分类:培养基配方] [发布时间:2021-8-5] [发布人:网站管理员2] [阅读次数:] [返回]

Rhodovulum iodosum
Rhodovulum robiginosum Medium
(DSMZ Medium 929)

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............................................................................................26.4g
MgSO 4 ·7H 2 O................................................................................6.8g
MgCl 2 ·6H 2 O..................................................................................5.7g
CaCl 2 ·2H 2 O...................................................................................1.5g
KCl..............................................................................................0.66g
KBr..............................................................................................0.09g
NaHCO 3 solution.....................................................................30.0mL
Phosphate solution...................................................................10.0mL
Sodium acetate solution...........................................................10.0mL
Iron sulfate solution.................................................................10.0mL
NH 4 Cl solution...........................................................................1.0mL
Thiosulfate solution ...................................................................1.0mL
Selenite-tungstate solution.........................................................1.0mL
Trace elements solution .............................................................1.0mL
Vitamin solution.........................................................................1.0mL
Vitamin B 12 solution..................................................................1.0mL
Vitamin B 1  solution....................................................................1.0mL
pH 6.8 ± 0.2 at 25°C
Sodium Acetate Solution:
Composition per 100.0mL:
Na-acetate .....................................................................................4.1g
Preparation of Sodium Acetate Solution: Add sodium acetate to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Filter sterilize.
Trace Elements Solution:
Composition per liter:
CoCl 2 ·6H 2 O ...........................................................................190.0mg
ZnSO 4 ·7H 2 O..........................................................................144.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
Na 2 MoO 4 ·4H 2 O .......................................................................36.0mg
H 3 BO 3 ......................................................................................30.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
Na 2 EDTA...................................................................................5.2mg
FeSO 4 ·7H 2 O...............................................................................2.1mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
Preparation of Trace Elements Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 6.0. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi
pressure–121°C. Cool to room temperature.
Vitamin Solution:
Composition per 100.0mL:
Pyridoxine................................................................................15.0mg
Nicotinate.................................................................................10.0mg
Pantothenate...............................................................................5.0mg
Para-aminobenzoic acid.............................................................4.0mg
Biotin.........................................................................................1.0mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
Vitamin B 12 Solution:
Composition per 100.0mL:
Cyanocobalamine ......................................................................5.0mg
Preparation of Vitamin B 12 Solution:  Add cyanocobalamine to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Filter sterilize.
Vitamin B 1 Solution:
Composition per 100.0mL:
Thiamine..................................................................................10.0mg
Preparation of Vitamin B 1 Solution:  Add thiamine to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
NH 4 Cl Solution:
Composition per 10.0mL:
NH 4 Cl ...........................................................................................2.5g
Preparation of NH 4 Cl Solution:  Add NH 4 Cl to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Thiosulfate Solution:
Composition per 10.0mL:
Na 2 S 2 O 3 ·5H 2 O............................................................................1.24g
Preparation of Thiosulfate Solution:  Add Na 2 S 2 O 3 ·5H 2 O to dis-
tilled/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.
Cool to room temperature.
NaHCO 3 Solution:
Composition per 100.0mL:
NaHCO 3 ........................................................................................8.4g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–
121°C under an atmosphere of CO 2 . Cool to room temperature.
Selenite-Tungstate Solution
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Selenite-Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Cool to room temperature.
Phosphate Solution:
Composition per 10.0mL:
KH 2 PO 4 .........................................................................................0.4g
Preparation of Phosphate Solution:  Add KH 2 PO 4 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Iron Sulfate Solution:
Composition per 10.0mL:
FeSO 4 ..........................................................................................1.52g
Preparation of Iron Sulfate Solution:  Add FeSO 4 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Preparation of Medium: Prepare and dispense medium under 90%
N 2 + 10% CO 2  gas mixture. Add components, except NaHCO 3 solu-
tion, phosphate solution, sodium acetate solution, iron sulfate solution,
NH 4 Cl solution, thiosulfate solution, selenite-tungstate solution, trace
elements solution, vitamin solution, vitamin B 12 solution, and vitamin
B 1  solution, to distilled/deionized water and bring volume to 933.0mL.
Mix thoroughly. Sparge with 90% N 2 + 10% CO 2  gas mixture. Auto-
clave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobi-
cally add 30.0mL NaHCO 3 solution, 10.0mL phosphate solution,
10.0mL sodium acetate solution, 10.0mL iron sulfate solution, 1.0mL
NH 4 Cl solution, 1.0mL thiosulfate solution, 1.0mL selenite-tungstate
solution, 1.0mL trace elements solution, 1.0mL vitamin solution,
1.0mL vitamin B 12 solution, and 1.0mL vitamin B 1  solution. When the
iron is added a white precipitate may form. Adjust pH to 6.8. Asepti-
cally and anaerobically distribute to tubes or bottles.
Use:  For the cultivation of Rhodovulum iodosum and Rhodovulum
robiginosum.