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培养基配方

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Thermotoga elfii Medium

[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]

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

Sodium thiosulfate·5H2O.............................................................. 5.0g

Pancreatic digest of casein............................................................ 5.0g

Yeast extract.................................................................................. 5.0g

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

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

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

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

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

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

Sodium acetate.............................................................................. 0.5g

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

Glucose solution ......................................................................20.0mL

L-Cysteine solution ..................................................................10.0mL

Na2CO3 solution......................................................................10.0mL

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

Wolfe’s mineral solution..........................................................10.0mL

Glucose Solution:

Composition per 20.0mL:

Glucose ......................................................................................... 4.0g

Preparation of Glucose Solution: Add glucose to distilled/deion

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

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

L-Cysteine Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O ..................................................................... 0.5g

Preparation of L-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.

Na2CO3 Solution:

Composition per 10.0mL:

Na2CO3 ......................................................................................... 2.0g

Preparation of Na2CO3 Solution: Add Na2CO3 to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–

121°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.

Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Before use, neutralize to pH 7.0 with sterile HCl.

Wolfe’s Mineral Solution:

Composition per liter:

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

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

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

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

CoCl2·6H2O .................................................................................. 0.1g

ZnSO4·7H2O ................................................................................. 0.1g

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

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

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

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

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

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

Na2MoO4·2H2O .......................................................................... 0.01g

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

Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with

KOH. Add remaining components. Add distilled/deionized water to

1.0L. Adjust pH to 6.8.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere. Add components, except glucose solu

tion, Na2CO3 solution, L-cysteine solution, and Na2S·9H2O solution,

to distilled/deionized water and bring volume to 950.0mL. Mix thor

oughly. Adjust pH to 6.8–7.0. Sparge with 80% N2 + 20% CO2. Auto

clave for 15 min at 15 psi pressure–121°C. Aseptically and

anaerobically add 20.0mL of sterile glucose solution, 10.0mL of sterile

Na2CO3 solution, 10.0mL of sterlie L-cysteine solution, and 10.0mL of

sterile Na2S·9H2O solution. Mix thoroughly. Aseptically and anaerobi

cally distribute into sterile tubes or bottles.

Use: For the cultivation of Thermotoga elfii.

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Thermotoga elfii Medium