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培养基配方

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Succiniclasticum Medium

[所属分类:培养基配方] [发布时间:2021-8-30] [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:

Disodium succinate....................................................................... 5.0g

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

NaCl ............................................................................................ 0.45g

(NH4)2SO4................................................................................... 0.45g

K2HPO4..................................................................................... 0.225g

KH2PO4..................................................................................... 0.225g

MgSO4·7H2O .............................................................................. 0.09g

CaCl2·2H2O................................................................................. 0.06g

Indigocarmine ............................................................................5.0mg

Rumen fluid, clarified ............................................................400.0mL

NaHCO3 solution .....................................................................10.0mL

L-Cysteine·HCl·H2O solution ..................................................10.0mL

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

pH: 6.6–6.8

NaHCO3 Solution:

Composition per 10.0mL:

NaHCO3 ........................................................................................ 6.4g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

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

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

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O ..................................................................... 0.3g

Preparation of L-Cysteine·HCl·H2O Solution: Add L-cyste

ine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL.

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

pressure–121°C.

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.

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

Before use, neutralize to pH 7.0 with sterile HCl.

Preparation of Medium: Add components, except Na2S·9H2O so

lution, L-cysteine·HCl·H2O solution, and NaHCO3 solution, to dis

tilled/deionized water and bring volume to 970.0mL. Mix thoroughly.

Gently heat and bring to boiling. Cool to room temperature while

sparging with 100% CO2. Autoclave for 15 min at 15 psi pressure–

121°C. Aseptically and anaerobically add 10.0mL of sterile L

cysteine·HCl·H2O solution, 10.0mL of sterile Na2S·9H2O solution, and

10.0mL of sterile NaHCO3 solution to each tube. Mix thoroughly.

Use: For the cultivation of Succiniclasticum ruminis.

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Succiniclasticum Medium