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TF(A) Medium (DSMZ Medium 740)

[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(A) Medium

(DSMZ Medium 740)

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

Composition per liter:

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

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

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

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

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

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

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

Calcium chloride solution........................................................10.0mL

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

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

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

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

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

pH 6.8 ± 0.2 at 25°C

Calcium Chloride Solution:

Composition per 10.0mL:

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

Preparation of Calcium Chloride Solution: Add CaCl2·2H2O

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

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

121°C. Cool to 25°C.

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 25°C.

Glucose Solution:

Composition per 10.0mL:

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. Filter sterilize.

Na2S·9H2O Solution:

Composition per 10.0mL:

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

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to distilled/

deionized water and bring volume to 10.0mL. Sparge with N2. Autoclave

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

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. Filter sterilize.

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: Prepare and dispense medium under an

oxygen-free 100% N2. Add components, except vitamin solution,

cysteine solution, calcium chloride solution, glucose solution, trace el

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

and bring volume to 940.0mL. Mix thoroughly. Sparge with 100% N2.

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

ly and anaerobically add 10.0mL sterile vitamin solution, 10.0mL of

sterile cysteine solution, 10.0mL sterile glucose solution, 10.0mL sterile

calcium chloride solution, 10.0mL sterile trace elements solution, and

10.0mL of sterile Na2S·9H2O solution. Mix thoroughly. Adjust pH to

6.8. Aseptically and anaerobically distribute into sterile tubes or flasks.

Use: For the cultivation of Fervidobacterium pennivorans, Fervido

bacterium gondwanense, and Fervidobacterium sp.

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TF(A) Medium (DSMZ Medium 740)