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XB45/XB90/PB90-2 Medium (DSMZ Medium 862)

[所属分类:培养基配方] [发布时间:2021-10-15] [发布人:网站管理员2] [阅读次数:] [返回]

XB45/XB90/PB90-2 Medium

(DSMZ Medium 862)

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

Composition per 1069.2mL:

Solution A ..............................................................................940.0mL

Solution E ..............................................................................100.0mL

Solution D ................................................................................10.0mL

Solution G ................................................................................10.0mL

Solution F...................................................................................7.2mL

Solution B ..................................................................................1.0mL

Solution C ..................................................................................1.0mL

pH 7.2 ± 0.2 at 25°C

Solution A:

Composition per 940.0mL:

NaCl .............................................................................................. 1.0g

KCl................................................................................................ 0.5g

MgCl2·6H2O.................................................................................. 0.4g

KH2PO4......................................................................................... 0.2g

NH4Cl ......................................................................................... 0.25g

CaCl2·2H2O................................................................................. 0.15g

Resazurin ...................................................................................0.5mg

Preparation of Solution A: Prepare under 80% N2 + 20% CO2 gas

atmosphere. Add components to distilled/deionized water and bring vol

ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N2

+ 20% CO2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

25°C.

Solution B:

Composition per liter:

FeCl2·4H2O ................................................................................... 1.5g

CoCl2·6H2O ...........................................................................190.0mg

MnCl2·4H2O...........................................................................100.0mg

ZnCl2........................................................................................70.0mg

Na2MoO4·2H2O .......................................................................36.0mg

NiCl2·6H2O ..............................................................................24.0mg

H3BO3 ........................................................................................6.0mg

CuCl2·2H2O ...............................................................................2.0mg

HCl (25% solution)..................................................................10.0mL

Preparation of Solution B: Add FeCl2·4H2O to 10.0mL of HCl so

lution. Mix thoroughly. Add distilled/deionized water and bring vol

ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with

80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–121°C.

Cool to 25°C.

Solution C:

Composition per liter:

Pyridoxine hydrochloride ......................................................300.0mg

Thiamine-HCl·2H2O..............................................................200.0mg

Nicotinic acid.........................................................................200.0mg

Vitamin B12 ............................................................................100.0mg

Calcium pantothenate ............................................................100.0mg

p-Aminobenzoic acid............................................................... 80.0mg

D(+)-Biotin............................................................................... 20.0mg

Preparation of Solution C: Add components to distilled/deionized

water and bring volume to 1.0L. Sparge with 100% N2. Mix thorough

ly. Filter sterilize.

Solution D:

Composition per liter:

Pyridoxine-HCl........................................................................ 10.0mg

Thiamine-HCl·2H2O.................................................................. 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 B12 ................................................................................ 0.1mg

Preparation of Solution D: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Sparge with 100%

N2. Filter sterilize.

Solution E:

Composition per 100.0mL:

NaHCO3........................................................................................ 5.0g

Preparation of Solution E: Add NaHCO3 to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Sparge with

100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

25°C.

Solution F:

Composition per 10.0mL:

Glucose ......................................................................................... 1.0g

Preparation of Solution F: Add glucose to distilled/deionized water

and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N2. Auto

clave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Solution G:

Composition per 10.0mL:

Na2S·9H2O................................................................................ 0.125g

Preparation of Solution G: Add Na2S·9H2O to distilled/deionized

water and bring volume to 10.0mL. Mix thoroughly. Autoclave under

100% N2 for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere. Sequentially add 1.0mL solution B,

1.0mL solution C, 10.0mL solution D, 100.0mL solution E, 7.2mL so

lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an

aerobically under 80% N2 + 20% CO2 into appropriate vessels. The pH

should be 7.2.

Use: For the cultivation of unclassified bacteria DSM 12558, DSM

12559, and DSM 12595.

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XB45/XB90/PB90-2 Medium (DSMZ Medium 862)