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Thermoanaerovibrio Medium (DSMZ Medium 873)

[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]

Thermoanaerovibrio Medium

(DSMZ Medium 873)

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

Composition per liter:

NaHCO3 ........................................................................................ 0.8g

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

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

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

CaCl2·2H2O................................................................................. 0.22g

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

Yeast extract................................................................................ 0.25g

Peptone........................................................................................ 0.25g

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

NaHCO3 solution .....................................................................20.0mL

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

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

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

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

Magnesium chloride solution...................................................10.0mL

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

pH 7.0–7.3 at 25°C

NaHCO3 Solution:

Composition per 20.0mL:

NaHCO3 ........................................................................................ 2.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 20.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2. Filter sterilize.

Magnesium Chloride Solution:

Composition per 10.0mL:

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

Preparation of Magnesium Chloride Solution: Add 0.33g of

MgCl2·6H2O to distilled/deionized water and bring volume to 10.0mL.

Mix thoroughly. Sparge with 100% N2. Filter sterilize.

Calcium Chloride Solution:

Composition per 10.0mL:

CaCl2·2H2O................................................................................. 0.22g

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. Filter sterilize.

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.................................................................................... 0.5g

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.

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.

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.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3 solu

tion, glucose solution, calcium chloride solution, magnesium chloride

solution, Na2S·9H2O solution, vitamin solution, and trace elements so

lution SL-10, to distilled/deionized water and bring volume to 929.0mL.

Mix thoroughly. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min

at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL

glucose solution, 10.0mL Na2S·9H2O solution, 10.0mL magnesium

chloride solution, 10.0ml calcium chloride solution, 10.0mL vitamin

solution, and 1.0mL trace elements solution SL-10. Mix thoroughly.

Adjust pH to 7.0–7.3 with 10.0mL NaHCO3 solution. Aseptically and

anaerobically distribute into sterile tubes or bottles.

Use: For the cultivation of Thermanaerovibrio velox (Thermosinus

velox).

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Thermoanaerovibrio Medium (DSMZ Medium 873)