Pseudomonas chloritidismutans Medium (DSMZ Medium 944)

[所属分类:培养基配方] [发布时间:2021-7-15] [发布人:网站管理员2] [阅读次数:] [返回]

Pseudomonas chloritidismutans Medium
(DSMZ Medium 944)

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

Composition per 1001.0mL:

Solution A..............................................................................900.0mL
Solution B................................................................................50.0mL
Solution C................................................................................50.0mL
Vitamin solution.........................................................................1.0mL
pH 9.0 ± 0.2 at 25°C
Solution A:
Composition per 900mL:
Na-acetate·3H 2 O.........................................................................2.72g
NaClO 3 ........................................................................................1.06g
KH 2 PO 4 .......................................................................................0.41g
Na 2 HPO 4 .....................................................................................0.53g
Resazurin...................................................................................0.5mg
Selenite-tungstate solution.........................................................4.0mL
Trace elements solution SL-10..................................................1.0mL
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 .
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 .
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Solution B:
Composition per 50.0mL:
CaCl 2 ...........................................................................................0.11g
MgCl 2 ..........................................................................................0.10g
Preparation of Solution B: Add components to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Sparge with 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room tem-
perature.
Solution C:
Composition per 50.0mL:
NaHCO 3 ......................................................................................3.73g
Na 2 S·9H 2 O....................................................................................0.5g
NH 4 HCO 3 ...................................................................................0.44g
Preparation of Solution C: Add components to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Sparge with 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room tem-
perature.
Vitamin Solution:
Composition per liter:
Vitamin B 12 ..............................................................................50.0mg
Pantothenic acid.......................................................................50.0mg
Riboflavin ................................................................................50.0mg
Alpha-lipoic acid......................................................................50.0mg
p-Aminobenzoic acid...............................................................50.0mg
Thiamine-HCl·2H 2 O................................................................50.0mg
Nicotinic acid...........................................................................25.0mg
Nicotine amide.........................................................................25.0mg
Biotin .......................................................................................20.0mg
Folic acid..................................................................................20.0mg
Pyridoxamine-HCl...................................................................10.0mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Prepare and dispense medium under
100% N 2 gas. Aseptically and anaerobically combine 900.0mL sterile
solution A, 50.0mL sterile solution B, 50.0mL sterile solution C, and
1.0mL sterile vitamin solution. Mix thoroughly. The pH should be 9.0.
Aseptically and anaerobically distribute into sterile tubes or flasks.
Use:  For the cultivation of Pseudomonas chloritidismutans (Pseudo-
monas stutzeri).