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Spirochete Thermophile Medium (DSMZ Medium 509)

[所属分类:培养基配方] [发布时间:2021-8-23] [发布人:网站管理员2] [阅读次数:] [返回]

Spirochete Thermophile Medium

(DSMZ Medium 509)

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

Composition per 1012.0mL:

Solution A..............................................................................920.0mL

Solution D................................................................................50.0mL

Solution E ................................................................................20.0mL

Solution F.................................................................................10.0mL

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

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

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

pH 7.0 ± 0.2 at 25°C

Solution A:

Composition per 920.0mL:

NaCl.............................................................................................. 4.0g

MgCl2·6H2O ................................................................................. 0.8g

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

NH4Cl ........................................................................................... 0.3g

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

CaCl2·2H2O ................................................................................ 0.03g

Resazurin ................................................................................... 1.0mg

Preparation of Solution A: Add components to 920.0mL distilled/

deionized water. Mix thoroughly. Bring to boiling for a few minutes.

Cool to room temperature while gassing with 80% N2 + 20% CO2 gas.

Adjust pH to 6.0. Immediately distribute under N2 into anaerobic tubes.

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:

NaOH............................................................................................ 0.5g

Na2WO4·2H2O........................................................................... 4.0mg

Na2SeO3·5H2O........................................................................... 3.0mg

Preparation of Solution C: Add components to distilled/deionized

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

Filter sterilize.

Solution D:

Composition per 100.0mL:

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

Preparation of Solution D: Add NaHCO3 to distilled/deionized

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

100% N2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C.

Cool to 25°C.

Solution E:

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 E: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% N2

+ 20% CO2. Filter sterilize.

Solution F:

Composition per 10.0mL:

Starch ............................................................................................ 1.0g

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

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

gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

25°C.

Solution G:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.3g

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: Solution A is distributed into anaerobic

tubes with rubber stoppers prior to autoclaving. Using aseptic and anaer

obic conditions and syringes, appropriate volumes of sterile solutions B–

G are injected into each tube to yield the specified concentrations.

Use: For the cultivation of Spirochaeta thermophila.

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Spirochete Thermophile Medium (DSMZ Medium 509)