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Spirochaeta smaragdinae Medium (DSMZ Medium 819)

[所属分类:培养基配方] [发布时间:2021-8-23] [发布人:网站管理员2] [阅读次数:] [返回]

Spirochaeta smaragdinae Medium

(DSMZ Medium 819)

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 ............................................................................................ 50.0g

Yeast extract.................................................................................. 5.0g

NH4Cl ........................................................................................... 1.0g

Cysteine-HCl·H2O ........................................................................ 0.5g

K2HPO4......................................................................................... 0.3g

KH2PO4......................................................................................... 0.3g

MgCl2·6H2O.................................................................................. 0.2g

KCl................................................................................................ 0.2g

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

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

NaHCO3 solution .....................................................................80.0mL

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

Trace elements solution ...........................................................10.0mL

pH 7.8 ± 0.2 at 25°C

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.2g

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.

Neutralize to pH 7.0 with sterile HCl.

NaHCO3 Solution:

Composition per 100.0mL:

NaHCO3 ........................................................................................ 5.0g

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.

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 Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3 solu

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

ume to 910.0mL. Mix thoroughly. Adjust pH to 7.8. Gently heat and

bring to boiling. Cool while sparging with 80% N2 + 20% CO2. Auto

clave for 15 min at 15 psi pressure–121°C. Cool to room temperature.

Aseptically and anaerobically add 80.0mL NaHCO3 solution and

10.0mL Na2S·9H2O solution. Aseptically and anaerobically distribute

into sterile tubes or bottles.

Use: For the cultivation of Spirochaeta smaragdinae.

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Spirochaeta smaragdinae Medium (DSMZ Medium 819)