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Thermoanaerobacter sulfurophilus Medium (DSMZ Medium 827)

[所属分类:培养基配方] [发布时间:2021-9-6] [发布人:网站管理员2] [阅读次数:] [返回]

Thermoanaerobacter sulfurophilus Medium

(DSMZ Medium 827)

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

Composition per 1055.0mL:

Sulfur, powdered......................................................................... 10.0g

NH4Cl ......................................................................................... 0.33g

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

KH2PO4....................................................................................... 0.33g

MgCl2·6H2O................................................................................ 0.33g

CaCl2·2H2O................................................................................. 0.33g

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

Glucose solution ......................................................................25.0mL

Na2S·9H2O solution .................................................................15.0mL

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

Yeast extract solution.................................................................5.0mL

Trace elements solution SL-10 ..................................................1.0mL

pH 7.0 ± 0.2 at 25°C

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 80% H2 + 20% CO2. Filter sterilize.

Glucose Solution:

Composition per 50.0mL:

Glucose ....................................................................................... 10.0g

Preparation of Glucose Solution: Add glucose to distilled/deion

ized water and bring volume to 50.0mL. Mix thoroughly. Filter steril

ize.

Yeast Extract Solution:

Composition per 10.0mL:

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

Preparation of Yeast Extract Solution: Add yeast extract 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.

Trace Elements Solution SL-10:

Composition per liter:

FeCl2·4H2O ................................................................................... 1.5g

CoCl2·6H2O ...........................................................................190.0mg

MnCl2·4H2O...........................................................................100.0mg

ZnCl2........................................................................................70.0mg

Na2MoO4·2H2O .......................................................................36.0mg

NiCl2·6H2O ..............................................................................24.0mg

H3BO3 ........................................................................................6.0mg

CuCl2·2H2O ...............................................................................2.0mg

HCl (25% solution)..................................................................10.0mL

Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O

to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized

water and bring volume to 1.0L. Add remaining components. Mix thor

oughly. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min at 15

psi pressure–121°C.

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. Mix thoroughly.

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

to room temperature.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas mixture. Add components, except Na2S·9H2O solu

tion, glucose solution, vitamin solution, and yeast extract solution, to

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

Heat to 90°C on each of 3 successive days. Aseptically and anaerobi

cally add 25.0mL sterile glucose solution, 15.0mL sterile Na2S·9H2O

solution, 10.0mL sterile vitamin solution, and 5.0mL sterile yeast ex

tract solution. Mix thoroughly. Aseptically and anaerobically distribute

to tubes or bottles. The pH should be 7.0.

Use: For the cultivation of Thermoanaerobacter sulfurophilus.

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Thermoanaerobacter sulfurophilus Medium (DSMZ Medium 827)