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Syntrophomonas species 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 1006.0mL:

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

Solution B ................................................................................70.0mL

Solution C ................................................................................10.0mL

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

pH 7.2 ± 0.2 at 25°C

Solution A:

Composition per 916.0mL:

Na2SO4.......................................................................................... 2.8g

Pancreatic digest of casein............................................................ 1.0g

Sodium stearate........................................................................... 0.61g

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

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

Rumen fluid, clarified..............................................................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. Adjust pH to 7.2. Gently heat and

bring to boiling. Continue boiling for a few minutes. Allow to cool to

room temperature under 80% N2 + 20% CO2. Distribute into bottles

under 80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–

121°C.

Mineral Solution:

Composition per liter:

Nitrilotriacetic acid ..................................................................... 12.5g

NaCl.............................................................................................. 1.0g

FeCl3·4H2O................................................................................... 0.2g

MnCl2·4H2O ................................................................................. 0.1g

CaCl2·2H2O .................................................................................. 0.1g

ZnCl2 ............................................................................................ 0.1g

CuCl2 .......................................................................................... 0.02g

Na2SeO3 ...................................................................................... 0.02g

CoCl2·6H2O.............................................................................. 0.017g

H3BO3......................................................................................... 0.01g

Na2MoO4·2H2O.......................................................................... 0.01g

Preparation of Mineral Solution: Add nitrilotriacetic acid to

500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add

remaining components. Add distilled/deionized water to 1.0L. Mix

thoroughly.

Vitamin Solution:

Composition per liter:

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

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

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

p-Aminobenzoic acid............................................................... 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.

Solution B:

Composition per 70.0mL:

NaHCO3 ........................................................................................ 3.5g

Preparation of Solution B: Add NaHCO3 to distilled/deionized

water and bring volume to 70.0mL. Mix thoroughly. Filter sterilize.

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

Solution C:

Composition per 10.0mL:

L-Cysteine·HCl.............................................................................. 0.3g

Preparation of Solution C: Add L-cysteine·HCl to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with

100% N2 for 3–4 min. Autoclave under 100% N2 for 15 min at 15 psi

pressure–121°C.

Solution D:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.3g

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

water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%

N2 for 3–4 min. Autoclave under 100% N2 for 15 min at 15 psi pres

sure–121°C.

Preparation of Medium: To 916.0mL of sterile solution A, add

70.0mL of sterile solution B, 10.0mL of sterile solution C, and 10.0mL

of sterile solution D. Mix thoroughly.

Use: For the cultivation of Syntrophomonas wolfei.

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Syntrophomonas species Medium