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Thermotoga hypogea Medium (DSMZ Medium 794)

[所属分类:培养基配方] [发布时间:2021-9-8] [发布人:网站管理员2] [阅读次数:] [返回]

Thermotoga hypogea Medium

(DSMZ Medium 794)

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

Composition per liter:

NaCl ............................................................................................ 10.0g

Yeast extract.................................................................................. 2.0g

Trypticase™.................................................................................. 2.0g

NH4Cl ........................................................................................... 1.0g

Na-acetate ..................................................................................... 0.5g

L-Cysteine ..................................................................................... 0.5g

K2HPO4......................................................................................... 0.3g

KH2PO4......................................................................................... 0.3g

MgCl2·6H2O.................................................................................. 0.2g

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

KCl................................................................................................ 0.1g

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

Na-thiosulfate solution.............................................................20.0mL

NaHCO3 solution .....................................................................20.0mL

Xylose solution ........................................................................20.0mL

Na2S·9H2O solution.................................................................14.0mL

Trace elements solution ...........................................................10.0mL

pH 7.3 ± 0.2 at 25°C

Xylose Solution:

Composition per 20.0mL:

Xylose........................................................................................... 3.0g

Preparation of Xylose Solution: Add xylose to distilled/deionized

water and bring volume to 20.0mL. Mix thoroughly. Sparge with 100%

N2. Filter sterilize.

NaHCO3 Solution:

Composition per 20.0mL:

NaHCO3........................................................................................ 2.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 20.0mL. Mix thoroughly. Autoclave

for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be prepared

freshly.

Na-thiosulfate Solution:

Composition per 20.0mL:

Na2S2O3·5H2O .............................................................................. 5.0g

Preparation of Na-thiosulfate Solution: Add Na2S2O3·5H2O to

distilled/deionized water and bring volume to 20.0mL. Mix thorough

ly. Sparge with 100% N2. Filter sterilize.

Na2S·9H2O Solution:

Composition per 20.0mL:

Na2S·9H2O.................................................................................... 0.6g

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

tilled/deionized water and bring volume to 20.0mL. Sparge with N2.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an

aerobically.

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 xylose solution,

Na2S·9H2O solution, Na-thiosulfate solution, and NaHCO3 solution, to

926.0mL distilled/deionized water. Mix thoroughly. Sparge with 80% N2

+ 20% CO2. Adjust pH to 7.2–7.4. Autoclave for 15 min at 15 psi pres

sure–121°C. Cool to 25°C while sparging with 80% N2 + 20% CO2.

Aseptically and anaerobically add 20.0mL sterile xylose solution,

14.0mL Na2S·9H2O solution, 20.0mL NaHCO3 solution, and 20.0mL

Na-thiosulfate solution, . Mix thoroughly. Aseptically and anaerobically

distribute into sterile tubes or flasks.

Use: For the cultivation of Thermotoga hypogea.

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Thermotoga hypogea Medium (DSMZ Medium 794)