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Thiohalophilus Medium (DSMZ Medium 1058)

[所属分类:培养基配方] [发布时间:2021-9-15] [发布人:网站管理员2] [阅读次数:] [返回]

Thiohalophilus Medium

(DSMZ Medium 1058)

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 ......................................................................................... 120.0g

K2HPO4......................................................................................... 1.5g

NH4Cl .......................................................................................... 0.5g

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

Calcium chloride solution........................................................10.0mL

Magnesium chloride solution...................................................10.0mL

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

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

Carbonate solution .................................................................. variable

pH 7.6 ± 0.2 at 25°C

Thiosulfate Solution:

Composition per 10.0mL:

Na2S2O3·5H2O .............................................................................. 5.0g

Preparation of Thiosulfate Solution: Add components to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Sparge with 100% N2. Filter sterilize.

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.

Carbonate Solution:

Composition per 10.0mL:

Na2CO3 ......................................................................................... 2.0g

Preparation of Carbonate Solution: Add Na2CO3 to distilled/de

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

with a gas mixture of 80% N2 + 20% CO2. Filter sterilize.

Magnesium Chloride Solution:

Composition per 10.0mL:

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

Preparation of Magnesium Chloride Solution: Add 0.4g of

MgCl2·6H2O to distilled/deionized water and bring volume to 10.0mL.

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

to room temperature. Sparge with 100% N2.

Calcium Chloride Solution:

Composition per 10.0mL:

CaCl2·2H2O ................................................................................ 0.05g

Preparation of Calcium Chloride Solution: Add components to

distilled/deionized water and bring volume to 10.0mL. Mix thorough

ly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room tem

perature. Sparge with 100% N2 gas.

Seven Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ...................................................................... 300.0mg

Nicotinic acid......................................................................... 200.0mg

Thiamine·HCl ........................................................................ 200.0mg

Calcium DL-pantothenate....................................................... 100.0mg

Cyanocobalamine .................................................................. 100.0mg

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

D(+)-Biotin............................................................................... 20.0mg

Preparation of Seven Vitamin Solution: Add components to dis

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

sterilize. Sparge with 100% N2.

Trace Elements Solution SL-10 with EDTA:

Composition per liter:

FeCl2·4H2O................................................................................... 1.5g

Na2-EDTA..................................................................................... 0.5g

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 with EDTA:

Add FeCl2·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis

tilled/deionized water and bring volume to 1.0L. Add remaining com

ponents. Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min

at 15 psi pressure–121°C. Adjust pH to 7.0.

Preparation of Medium: Add components, except magnesium

chloride, thiosulfate, bicarbonate, and vitamin solutions, to distilled/

deionized water and bring volume to 970.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2 gas mixture for at least 30 min to remove dis

solved oxygen and to saturate the solution with CO2. Dispense into an

aerobic culture vessels (e.g., Balch tubes) to 1/2 volume under air

atmosphere. Close vials with butyl rubber septa to prevent free ex

change of oxygen with the external atmosphere. Autoclave for 15 min

at 15 psi pressure–121°C. Cool to room temperature. Aseptically add

magnesium chloride, thiosulfate, bicarbonate, and vitamin solutions.

Mix thoroughly. Adjust pH to 7.5–7.8 using a sterile solution of sodium

carbonate (5% w/v).

Use: For the cultivation of Thiohalomonas denitrificans, Thiohalo

philus thiocyanatoxydans, and Thiomicrospira halophila.

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Thiohalophilus Medium (DSMZ Medium 1058)