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Thermococcus Medium (DSMZ Medium 806)

[所属分类:培养基配方] [发布时间:2021-9-7] [发布人:网站管理员2] [阅读次数:] [返回]

Thermococcus Medium

(DSMZ Medium 806)

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 ............................................................................................ 18.0g

Sulfur ............................................................................................ 5.0g

MgSO4·7H2O ................................................................................ 3.4g

MgCl2·2H2O.................................................................................. 2.7g

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

Trypticase™.................................................................................. 1.0g

NaHCO3 ........................................................................................ 1.0g

KCl.............................................................................................. 0.33g

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

CaCl2·2H2O................................................................................. 0.14g

K2HPO4....................................................................................... 0.14g

Resazurin .................................................................................. 0.001g

Na2SeO3 .................................................................................0.001mg

NiCl2·6H2O ............................................................................0.001mg

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

Vitamin solution.......................................................................10.0mL

Cysteine solution......................................................................10.0mL

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

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. Adjust pH to 7.0.

Cysteine Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O ..................................................................... 0.3g

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O to

distilled/deionized water and bring volume to 10.0mL. Mix thorough

ly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to room temperature.

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.

Vitamin Solution:

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 Vitamin Solution: Add components to distilled/

deionized water and bring volume to 1.0L. Mix thoroughly. Sparge

with 100% N2. Filter sterilize.

Preparation of Medium: Add components, except Fildes enrich

ment solution, NaHCO3, vitamin solution, cysteine solution, and

Na2S·9H2O solution, to distilled/deionized water and bring volume to

970.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 5

min. Cool to room temperature under 100% N2. Add 1.0g solid

NaHCO3. Adjust pH to 7.2. Sterilize at 100°C for 3 hr on 3 consecutive

days. Aseptically and anaerobically add 10.0mL vitamin solution,

10.0mL cysteine solution, and 10.0mL Na2S·9H2O solution. Mix thor

oughly. Adjust pH to 7.2. Aseptically and anaerobically distribute to ster

ile tubes or bottles.

Use: For the cultivation of Thermococcus spp.

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Thermococcus Medium (DSMZ Medium 806)