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SRB-Psychrophile Medium (DSMZ Medium 861)

[所属分类:培养基配方] [发布时间:2021-8-24] [发布人:网站管理员2] [阅读次数:] [返回]

SRB-Psychrophile Medium

(DSMZ Medium 861)

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

Composition per 1158mL:

NaCl............................................................................................ 20.0g

Na2SO4.......................................................................................... 4.0g

MgCl2·6H2O ................................................................................. 3.0g

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

KBr ............................................................................................. 0.09g

KCl................................................................................................ 0.5g

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

NH4Cl solution ........................................................................49.5mL

KH2PO4 solution......................................................................49.5mL

NaHCO3 solution .....................................................................29.7mL

Vitamin solution.........................................................................9.9mL

Substrate solution.......................................................................9.9mL

Na2S·9H2O solution...................................................................9.9mL

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

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

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

pH 7.2 ± 0.2 at 25°C

Dithionite Solution

Composition per 10.0mL:

Na-dithionite............................................................................... 0.25g

Preparation of Dithionite Solution: Add Na-dithionite to dis

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

Sparge with 100% N2. Filter sterilize.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.5g

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.

Vitamin Solution:

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 Vitamin Solution: Add components to distilled/

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

with 80% H2 + 20% CO2. Filter sterilize.

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.

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.

NaHCO3 Solution:

Composition per 100.0mL:

NaHCO3 ...................................................................................... 10.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2. Filter sterilize.

NH4Cl Solution:

Composition per 100.0mL:

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

Preparation of NH4Cl Solution: Add NH4Cl to distilled/deion

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

100% N2. Filter sterilize.

KH2PO4 Solution:

Composition per 100.0mL:

KH2PO4......................................................................................... 0.4g

Preparation of KH2PO4 Solution: Add KH2PO4 to distilled/de

ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge

with 100% N2. Filter sterilize.

Substrate Solution:

Composition per 10.0mL:

Na-acetate ..................................................................................... 1.5g

Preparation of Substrate Solution: Add Na-acetate to distilled/de

ionized water and bring volume to 10.0mL. Sparge with N2. Filter sterilize.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2. Add components, except NaHCO3 solution,

Na2S·9H2O solution, substrate solution, selenite-tungstate solution,

NH4Cl solution, KH2PO4 solution, vitamin solution, dithionite solu

tion, and trace elements solution SL-10. Distribute 30.0mL aliquots

into 50mL serum bottles. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to room temperature. Aseptically and anaerobically for

each 30.0mL medium add 1.5mL NH4Cl solution, 1.5mL KH2PO4 so

lution, 0.9mL NaHCO3 solution, and 0.3mL each of Na2S·9H2O, sub

strate, selenite-tungstate, and vitamin solutiona, and 0.03mL each of

dithionite solutionand trace elements solution SL-10. Final pH is 7.2.

Use: For the cultivation of Desulfofrigus oceanense DSM 12341.

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SRB-Psychrophile Medium (DSMZ Medium 861)