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ZF2 Medium (DSMZ Medium 943)

[所属分类:培养基配方] [发布时间:2021-10-25] [发布人:网站管理员2] [阅读次数:] [返回]

ZF2 Medium

(DSMZ Medium 943)

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

Composition per liter:

NaHCO3 ........................................................................................ 3.8g

Yeast extract.................................................................................. 3.0g

(NH4)HCO3................................................................................. 0.45g

MgSO4·6H2O .............................................................................. 0.13g

CaCl2·2H2O................................................................................. 0.12g

Resazurin ...................................................................................0.5mg

Phosphate buffer ......................................................................10.0mL

Glycine solution.........................................................................5.0mL

Arginine solution .......................................................................5.0mL

Na2S·9H2O solution ...................................................................5.0mL

Dithionite solution .....................................................................1.0mL

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

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

Selenite-tungstate solution.........................................................1.0mL

pH 7.3 ± 0.2 at 25°C

Arginine Solution:

Composition per 10.0mL:

Arginine-HCl ................................................................................ 3.5g

Preparation of Arginine Solution: Add arginine-HCl to distilled/

deionized water and bring volume to 10.0mL. Mix thoroughly. Filter

sterilize.

Glycine Solution:

Composition per 100.0mL:

Glycine........................................................................................ 15.0g

Preparation of Glycine Solution: Add glycine to distilled/deion

ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with

100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

room temperature.

Na2S·9H2O Solution:

Composition per 10.0mL:

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

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

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

Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool

to room temperature.

Dithionite Solution:

Composition per 10.0mL:

Na2-dithionite.............................................................................. 0.25g

Preparation of Dithionite Solution: Add Na2-dithionite to distilled/

deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave

under 100% N2 for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Seven Vitamin Solution:

Composition per liter:

Pyridoxine hydrochloride ...................................................... 300.0mg

Thiamine-HCl·2H2O.............................................................. 200.0mg

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

Vitamin B12 ............................................................................ 100.0mg

Calcium pantothenate ............................................................ 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. Sparge with 100% N2.

Mix thoroughly. Filter sterilize.

Phosphate Buffer:

Composition per liter:

Na2HPO4·12H2O......................................................................... 43.0g

KH2PO4....................................................................................... 5.44g

Preparation of Phosphate Buffer: Add components to distilled/

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

to 7.3. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°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: 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. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min at 15

psi pressure–121°C.

Selenite-Tungstate Solution

Composition per liter:

NaOH............................................................................................ 0.5g

Na2WO4·2H2O........................................................................... 4.0mg

Na2SeO3·5H2O........................................................................... 3.0mg

Preparation of Selenite-Tungstate Solution: Add components

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

Sparge with 100% N2. Filter sterilize.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas mixture. Add components, except phosphate buffer,

glycine solution, arginine solution, dithionite solution, seven vitamin so

lution, and Na2S·9H2O solution, to distilled/deionized water and bring

volume to 973.0mL. Mix thoroughly. Equilibrate with 80% N2 + 20%

CO2 to reach pH 7.3. Autoclave for 15 min at 15 psi pressure–121°C.

Cool to 25°C. Aseptically and anaerobically add 10.0mL phosphate buf

fer, 5.0mL glycine solution, 5.0mL arginine solution, 1.0mL dithionite

solution, 1.0mL seven vitamin solution, and 5.0mL Na2S·9H2O solution.

Mix thoroughly. Aseptically and anaerobically distribute into sterile

tubes or flasks.

Use: For the cultivation of Sedimentibacter saalensis.

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ZF2 Medium (DSMZ Medium 943)