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Stetteria Medium (DSMZ Medium 795)

[所属分类:培养基配方] [发布时间:2021-8-26] [发布人:网站管理员2] [阅读次数:] [返回]

Stetteria Medium

(DSMZ Medium 795)

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

Composition per liter:

Sulfur, powdered......................................................................... 10.0g

Peptone.......................................................................................... 2.0g

Yeast extract.................................................................................. 1.0g

KH2PO4......................................................................................... 0.5g

NaHCO3 ...................................................................................... 0.16g

NiCl2·6H2O ................................................................................3.0mg

Resazurin .................................................................................0.75mg

Synthetic seawater, concentrated ...........................................500.0mL

Trace elements solution ...........................................................15.0mL

Na2S·9H2O solution .................................................................10.0mL

Selenite-tungstate solution.........................................................1.5mL

pH 7.2 ± 0.2 at 25°C

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.5g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

tilled/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 room temperature.

Selenite-Tungstate Solution

Composition per liter:

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

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

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

Preparation of Selenite-Tungstate Solution: Add components

to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Sparge with 100% N2.

Synthetic Seawater, Concentrated:

Composition per liter:

NaCl ............................................................................................ 55.4g

MgSO4·7H2O .............................................................................. 14.0g

MgCl2·6H2O................................................................................ 11.0g

CaCl2·2H2O................................................................................... 1.5g

KCl................................................................................................ 1.3g

NaBr.............................................................................................. 0.2g

H3BO3 ......................................................................................... 0.06g

SrCl2·6H2O.................................................................................. 0.03g

Na3-citrate ................................................................................20.0mg

KI .............................................................................................. 0.1mg

Preparation of Synthetic Seawater, Concentrated: Add com

ponents to distilled/deionized water and bring volume to 1.0L. Mix

thoroughly. Filter sterilize.

Trace Elements Solution:

Composition per liter:

MgSO4·7H2O ................................................................................ 3.0g

Nitrilotriacetic acid ....................................................................... 1.5g

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

MnSO4·2H2O................................................................................ 0.5g

CoSO4·7H2O............................................................................... 0.18g

ZnSO4·7H2O ............................................................................... 0.18g

CaCl2·2H2O .................................................................................. 0.1g

FeSO4·7H2O.................................................................................. 0.1g

NiCl2·6H2O............................................................................... 0.025g

KAl(SO4)2·12H2O....................................................................... 0.02g

H3BO3 ......................................................................................... 0.01g

Na2MoO4·4H2O .......................................................................... 0.01g

CuSO4·5H2O............................................................................... 0.01g

Na2SeO3·5H2O........................................................................... 0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH

to 6.5 with KOH. Add remaining components. Add distilled/deionized

water to 1.0L. Mix thoroughly.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas mixture. Add components, except synthetic seawa

ter and Na2S·9H2O solution, to 490.0mL distilled/deionized water. Mix

thoroughly. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min at

15 psi pressure–121°C. Cool to 25°C. Aseptically add 500.0mL filter

sterilized concentrated seawater. Flush with 80% N2 + 20% CO2 gas

mixture for 20 min. Aseptically add 10.0mL Na2S·9H2O solution. Ad

just pH to 6.0 with H2SO4. Mix thoroughly. Aseptically and anaerobi

cally distribute 20mL aliquots into sterile 100mL serum bottles.

Pressurize bottles to 2 bar gas overpressure with 80% N2 + 20% CO2.

Heat at 100°C for 1.5 h. Before use check that the medium pH is 6.0.

Use: For the cultivation of Stetteria hydrogenophila and Staphylother

mus hellenicus.

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Stetteria Medium (DSMZ Medium 795)