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培养基配方

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Thermodesulforhabdus Medium

[所属分类:培养基配方] [发布时间:2021-9-7] [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 liter:

NaCl............................................................................................ 10.0g

Na2SO4.......................................................................................... 7.0g

Sodium acetate·3H2O.................................................................... 6.2g

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

KH2PO4......................................................................................... 1.0g

NH4Cl ......................................................................................... 0.25g

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

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

Na2S·9H2O solution .................................................................10.0mL

Na2S2O4 solution......................................................................10.0mL

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

NaHCO3 solution .................................................................... variable

pH 6.8 ± 0.2 at 25°C

NaHCO3 Solution:

Composition per 20.0mL:

NaHCO3 ........................................................................................ 1.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 20.0mL. Mix thoroughly. Sparge

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

121°C.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................. 0.15g

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

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

Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Na2S2O3 Solution:

Composition per 10.0mL:

Na2S2O3·5H2O .............................................................................. 2.0g

Preparation of Na2S2O3 Solution: Add Na2S2O3·5H2O to dis

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

Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–121°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.

Preparation of Medium: Prepare and dispense medium under

100% N2. Add components, except Na2S·9H2O solution, Na2S2O3 solu

tion, and NaHCO3 solution, to distilled/deionized water and bring vol

ume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Cool

to room temperature while sparging with 100% N2. Anaerobically dis

tribute 9.8mL volumes into tubes. Autoclave for 15 min at 15 psi pres

sure–121°C. Aseptically and anaerobically add 0.1mL of sterile

Na2S·9H2O solution and 0.1mL of sterile Na2S2O4 solution to each tube.

Aseptically and anaerobically add a sufficient volume of sterile NaHCO3

solution to each tube to bring the pH to 6.8.

Use: For the cultivation of Thermodesulforhabdus norvegicus.

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Thermodesulforhabdus Medium