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Syntrophobacter pfennigii Medium

[所属分类:培养基配方] [发布时间:2021-9-1] [发布人:网站管理员2] [阅读次数:] [返回]

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.............................................................................................. 1.0g

Na2SO4.......................................................................................... 0.7g

KCl................................................................................................ 0.5g

MgCl2·6H2O ................................................................................. 0.4g

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

KH2PO4......................................................................................... 0.2g

CaCl2·2H2O ................................................................................ 0.15g

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

Sodium propionate solution.....................................................50.0mL

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

Na2S2O4 solution .....................................................................10.0mL

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

Seven vitamin solution ..............................................................1.0mL

NaHCO3 solution.................................................................... variable

pH 7.2–7.4 at 25°C

Sodium Propionate Solution:

Composition per 50.0mL:

Sodium propionate........................................................................ 1.5g

Preparation of Sodium Propionate Solution: Add sodium pro

pionate to distilled/deionized water and bring volume to 50.0mL. Mix

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

NaHCO3 Solution:

Composition per 20.0mL:

NaHCO3........................................................................................ 2.5g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 20.0mL. Mix thoroughly. Filter ster

ilize. Sparge with 80% N2 + 20% CO2.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................. 0.36g

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.

Na2S2O4 Solution:

Composition per 10.0mL:

Na2S2O4·5H2O .............................................................................. 2.0g

Preparation of Na2S2O4 Solution: Add Na2S2O4·5H2O 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.

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: Prepare and dis

pense under 80% N2 + 20% CO2. Add FeCl2·4H2O to 10.0mL of HCl

solution. Mix thoroughly. Add distilled/deionized water and bring vol

ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with

80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–121°C.

Seven Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ............................................................................. 0.3g

Thiamine·HCl ............................................................................... 0.2g

Nicotinic acid................................................................................ 0.2g

Calcium DL-pantothenate.............................................................. 0.1g

Vitamin B12 ................................................................................... 0.1g

p-Aminobenzoic acid...............................................................80.0mg

Biotin .......................................................................................20.0mg

Preparation of Seven Vitamin Solution: Add components to

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

Preparation of Medium: Prepare medium and dispense under 80%

N2 + 20% CO2. Add components, except sodium propionate solution,

NaHCO3 solution, Na2S·9H2O solution, and Na2S2O3 solution, to dis

tilled/deionized water and bring volume to 930.0mL. Mix thoroughly.

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

sure–121°C. Aseptically and anaerobically add 50.0mL of sterile sodi

um propionate solution, 10.0mL of sterile Na2S2O3 solution, and

10.0mL of sterile Na2S·9H2O solution. Mix thoroughly. Aseptically

and anaerobically add sufficient volume of sterile NaHCO3 solution to

bring pH to 7.2–7.4. Aseptically and anaerobically distribute into ster

ile tubes or flasks.

Use: For the cultivation of Syntrophobacter pfennigii.

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Syntrophobacter pfennigii Medium