Mineral Medium with Quinoline (DSMZ Medium 461a)

[所属分类:培养基配方] [发布时间:2021-6-2] [发布人:网站管理员2] [阅读次数:] [返回]

Mineral Medium with Quinoline
(DSMZ Medium 461a)

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

Composition per liter:

MgSO 4 ·7H 2 O................................................................................0.5g
NH 4 Cl ...........................................................................................0.3g
NaCl............................................................................................0.05g
CaCl 2 ..........................................................................................0.01g
MnSO 4 ........................................................................................0.01g
Phosphate solution.................................................................100.0mL
Quinoline emulsion................................................................100.0mL
Vitamin solution.........................................................................5.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
H 3 BO 3 ....................................................................................300.0mg
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
CuCl 2 ·2H 2 O ...............................................................................2.0mg
HCl (25% solution)....................................................................7.7mL
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 compo-
nents. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at
15 psi pressure–121°C. Cool to room temperature.
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
Nicotinamide............................................................................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.
Phosphate Solution:
Composition per 100.0mL:
Na 2 HPO 4 ·2H 2 O...........................................................................1.45g
KH 2 PO 4 .......................................................................................0.25g
Preparation of Phosphate Solution:  Add components to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper-
ature.
Quinoline Emulsion:
Composition per liter:
Quinoline ......................................................................................3.0g
Preparation of Quinoline Emulsion:  Prepare an emulsion of qui-
noline in 50 mM phosphate buffer by stirring or ultrasonication. Add
quinoline acid to phosphate buffer (50 mM, pH 7.5) and bring volume
to 1.0L. Stir vigorously or sonicate to form emulsion. Autoclave for 15
min at 15 psi pressure–121°C in a gas tight vessel. Cool to room tem-
perature.
Preparation of Medium:  Add components, except phosphate solu-
tion, vitamin solution, and quinoline emulsion to distilled/deionized
water and bring volume to 795.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to room temperature. Aseptically
add 100.0mL phosphate solution, 5.0mL vitamin solution, and
100.0mL quinoline emulsion. Mix thoroughly. Aseptically distribute
into sterile tubes or flasks
Use: For the cultivation of Rhodococcus rhodochrous (Rhodococcus
roseus) and other quinoline-utilizing bacteria.