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培养基配方

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TF Medium

[所属分类:培养基配方] [发布时间:2021-9-3] [发布人:网站管理员2] [阅读次数:] [返回]

TF Medium

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.............................................................................................. 7.0g

Pancreatic digest of casein............................................................ 2.0g

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

MgSO4·7H2O................................................................................ 1.8g

K2HPO4......................................................................................... 1.6g

MgCl2·6H2O ................................................................................. 1.4g

Na2HPO4·H2O............................................................................... 1.0g

NH4Cl ........................................................................................... 0.5g

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

MgSO4·7H2O.............................................................................. 0.16g

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

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

Wolfe’s vitamin solution..........................................................10.0mL

Glucose solution ......................................................................10.0mL

CaCl2·2H2O solution................................................................10.0mL

L-Cysteine·HCl·H2O solution ..................................................10.0mL

pH 6.8–7.0 at 25°C

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.

Wolfe’s Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ........................................................................10.0mg

p-Aminobenzoic acid.................................................................5.0mg

Lipoic acid .................................................................................5.0mg

Nicotinic acid.............................................................................5.0mg

Riboflavin ..................................................................................5.0mg

Thiamine·HCl ............................................................................5.0mg

Calcium DL-pantothenate...........................................................5.0mg

Biotin .........................................................................................2.0mg

Folic acid....................................................................................2.0mg

Vitamin B12 ................................................................................0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to

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

Filter sterilize.

Glucose Solution:

Composition per 10.0mL:

D-Glucose...................................................................................... 3.0g

Preparation of Glucose Solution: Add glucose to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with

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

CaCl2·2H2O Solution:

Composition per 10.0mL:

CaCl2·2H2O................................................................................. 0.06g

Preparation of CaCl2·2H2O Solution: Add CaCl2·2H2O 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.

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

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

Preparation of L-Cysteine·HCl·H2O Solution: Add L-cyste

ine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL.

Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi

pressure–121°C.

Preparation of Medium: Prepare and dispense medium under

100% N2. Add components, except glucose solution, CaCl2·2H2O so

lution, and L-cysteine·HCl·H2O solution, to distilled/deionized water

and bring volume to 970.0mL. Mix thoroughly. Adjust pH to 6.8–7.0.

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

Aseptically and anaerobically add 10.0mL of sterile glucose solution,

10.0mL of sterile CaCl2·2H2O solution, and 10.0mL of sterile L

cysteine·HCl·H2O solution. Mix thoroughly. Final pH should be 6.8–

7.0.

Use: For the cultivation of Thermosipho species.

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TF Medium