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Thiofaba Medium (DSMZ Medium 1114)

[所属分类:培养基配方] [发布时间:2021-9-14] [发布人:网站管理员2] [阅读次数:] [返回]

Thiofaba Medium

(DSMZ Medium 1114)

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

Composition per liter:

Solution A..............................................................................960.0mL

Solution B ................................................................................10.0mL

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

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

Solution E ................................................................................10.0mL

pH 6.5 ± 0.2 at 25°C

Solution A:

Composition per 960.0mL:

MgCl2·6H2O................................................................................ 0.75g

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

NH4Cl ........................................................................................ 0.54g

Trace elements solution .............................................................2.0mL

Trace Elements Solution:

Composition per liter:

MgSO4·7H2O ................................................................................ 3.0g

Nitrilotriacetic acid ....................................................................... 1.5g

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

MnSO4·2H2O ................................................................................ 0.5g

CoSO4·7H2O ............................................................................... 0.18g

ZnSO4·7H2O ............................................................................... 0.18g

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

FeSO4·7H2O.................................................................................. 0.1g

NiCl2·6H2O ............................................................................... 0.025g

KAl(SO4)2·12H2O....................................................................... 0.02g

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

Na2MoO4·4H2O .......................................................................... 0.01g

CuSO4·5H2O ............................................................................... 0.01g

Na2SeO3·5H2O...........................................................................0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH

to 6.5 with KOH. Add remaining components. Add distilled/deionized

water to 1.0L. Mix thoroughly.

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 960.0mL. Mix thoroughly. Sparge with a

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

pressure–121°C. Cool to room temperature.

Solution B:

Composition per 10.0mL:

KH2PO4....................................................................................... 1.19g

KH2PO4....................................................................................... 0.21g

Preparation of Solution B: Add components to distilled/deionized

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

N2. Autoclave or 15 min at 15 psi pressure–121°C. Cool to room tem

perature.

Solution C:

Composition per 10.0mL:

Na2CO3 ......................................................................................... 4.0g

Preparation of Solution C: Add Na2CO3 to distilled/deionized wa

ter and bring volume to 10.0mL. Mix thoroughly. Sparge with a gas

mixture of 80% N2 + 20% CO2. Filter sterilize.

Solution D:

Composition per 10.0mL:

Na2S2O3·5H2O ............................................................................ 1.25g

Preparation of Solution D: Add Na2S2O3·5H2O to distilled/deion

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

100% N2. Filter sterilize.

Solution E:

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 Solution E: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Sparge with 100%

N2. Filter sterilize.

Preparation of Medium: Dispense solution A under a gas atmo

sphere of 80% N2 + 20% CO2 into serum vials to 20% of volume. Au

toclave for 15 min at 15 psi pressure–121°C. Cool to room tempera

ture. Aspetically add sterile solutions B–E in that order. Adjust the pH

to 6.5. After inoculation pressurize with an amount of sterile air that is

equivalent to 30% of the volume of the cultivation vessel.

Use: For the cultivation of Thiofaba spp.

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Thiofaba Medium (DSMZ Medium 1114)