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Tepidibacter Medium with Peptone (DSMZ Medium 985)

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

Tepidibacter Medium with Peptone

(DSMZ Medium 985)

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 ........................................................................................... 18.0g

Peptone........................................................................................ 10.0g

MgCl2·6H2O.................................................................................. 4.0g

KCl ............................................................................................. 0.34g

NH4Cl ........................................................................................ 0.25g

Yeast extract ................................................................................. 0.2g

KH2PO4....................................................................................... 0.18g

CaCl2·2H2O................................................................................. 0.11g

Fe(NH4)2(SO4)2·6H2O ............................................................... 0.02g

Resazurin ..................................................................................1.0mg

Bicarbonate solution ................................................................10.0mL

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

Sulfide solution ........................................................................10.0mL

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

Wolfe's mineral elixir.................................................................1.0mL

pH 6.8 ± 0.2 at 25°C

Sulfide Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.5g

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

ionized 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.

Bicarbonate Solution:

Composition per 10.0mL:

NaHCO3 ........................................................................................ 5.0g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

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

with a gas mixture of 80% N2 + 20% CO2. 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 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 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C.

Wolfe’s Mineral Elixir:

Composition per liter:

MgSO4·7H2O.............................................................................. 30.0g

NaCl............................................................................................ 10.0g

MnSO4·2H2O................................................................................ 5.0g

(NH4)2NiSO4·6H2O ...................................................................... 2.8g

CoCl2·6H2O .................................................................................. 1.8g

ZnSO4·7H2O ................................................................................. 1.8g

FeSO4·7H2O.................................................................................. 1.0g

CaCl2·2H2O .................................................................................. 1.0g

KAl(SO4)2·12H2O....................................................................... 0.18g

CuSO4·5H2O................................................................................. 0.1g

H3BO3 ........................................................................................... 0.1g

Na2MoO4·2H2O ............................................................................ 0.1g

Na2SeO4 ........................................................................................ 0.1g

Na2WO4·2H2O.............................................................................. 0.1g

Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of dis

tilled/deionized water to 1.0 with dilute H2SO4. Add remaining com

ponents one at a time. Mix throughly to dissolve.

Preparation of Medium: Add components, except vitamin, bicar

bonate, and sulfide solutions, to distilled/deionized water and bring

volume to 970.0mL. Mix thoroughly. Gently heat and bring to boiling.

Boil for 1 min. Cool to room temperature while sparging with a gas

mixture of 80% N2 + 20% CO2. Dispense into culture vessels under an

atmosphere of 80% N2 + 20% CO2. Autoclave for 15 min at 15 psi

pressure–121°C. Cool to room temperature. Aspetically add vitamin,

bicarbonate, and sulfide solutions. Mix thoroughly. Adjust the pH to

6.6–7.0.

Use: For the cultivation of Clostridium tepidiprofundi.

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Tepidibacter Medium with Peptone (DSMZ Medium 985)