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Syntrophococcus sucromutans 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 1002.0mL:

Solution A ..............................................................................916.0mL

Solution C ................................................................................50.0mL

Solution B ................................................................................25.0mL

Solution D ................................................................................10.0mL

Solution E ..................................................................................1.0mL

pH 7.2–7.4 at 25°C

Solution A:

Composition per 916.0mL:

Sodium formate............................................................................. 0.6g

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

Rumen fluid, clarified ............................................................300.0mL

Mineral solution.......................................................................50.0mL

Vitamin solution.........................................................................5.0mL

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

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 916.0mL. Mix thoroughly. Adjust pH to 6.4.

Autoclave for 15 min at 15 psi pressure–121°C.

Mineral Solution:

Composition per liter:

KH2PO4....................................................................................... 10.0g

NaCl .............................................................................................. 8.0g

NH4Cl ........................................................................................... 8.0g

MgCl2·6H2O.................................................................................. 6.6g

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

Preparation of Mineral Solution: Add components to distilled/

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

Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ..........................................................................6.2mg

Nicotinic acid.............................................................................2.5mg

p-Aminobenzoic acid............................................................... 1.25mg

Thiamine·HCl .......................................................................... 1.25mg

Pantothenic acid....................................................................... 0.62mg

Biotin ....................................................................................... 0.25mg

Preparation of Vitamin Solution: Add components to distilled/

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

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. Autoclave for 15 min at 15 psi pressure–121°C.

Solution B:

Composition per 25.0mL:

Lactose.......................................................................................... 5.0g

Preparation of Solution B: Add lactose to distilled/deionized wa

ter and bring volume to 25.0mL. Mix thoroughly. Filter sterilize.

Solution C:

Composition per 50.0mL:

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

Preparation of Solution C: Add NaHCO3 to distilled/deionized

water and bring volume to 50.0mL. Mix thoroughly. Autoclave for 15

min at 15 psi pressure–121°C.

Solution D:

Composition per 10.0mL:

L-Cysteine ................................................................................... 0.24g

Preparation of Solution D: Add L-cysteine to distilled/deionized

water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15

min at 15 psi pressure–121°C.

Solution E:

Composition per 1.0mL:

Na2S·9H2O............................................................................... 78.0mg

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

water and bring volume to 1.0mL. Mix thoroughly. Autoclave for 15

min at 15 psi pressure–121°C.

Preparation of Medium: Prepare and dispense medium under H2-

free 80% N2 + 20% CO2. Aseptically and anaerobically combine

916.0mL of sterile solution A with 25.0mL of sterile solution B,

50.0mL of sterile solution C, 10.0mL of sterile solution D, and 1.0mL

of sterile solution E. Mix thoroughly. Final pH should be 6.4–6.8

Use: For the cultivation and maintenance of Syntrophococcus sucro

mutans.

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Syntrophococcus sucromutans Medium