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Sporomusa Medium

[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]

Sporomusa Medium

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

Composition per 1010.0mL:

Betaine·H2O .................................................................................. 6.7g

NaHCO3 ........................................................................................ 4.0g

NaCl ............................................................................................ 2.25g

Pancreatic digest of casein............................................................ 2.0g

Yeast extract.................................................................................. 2.0g

MgSO4·7H2O ............................................................................... 0.5g

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

K2HPO4..................................................................................... 0.348g

CaCl2·2H2O................................................................................. 0.25g

KH2PO4..................................................................................... 0.227g

FeSO4·7H2O...............................................................................2.0mg

Resazurin ...................................................................................1.0mg

NaHSeO3...................................................................................26.3μg

Vitamin solution.......................................................................10.0mL

Reducing agent solution...........................................................10.0mL

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

pH 7.0 ± 0.2 at 25°C

Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ........................................................................10.0mg

Calcium DL-pantothenate...........................................................5.0mg

Lipoic acid .................................................................................5.0mg

Nicotinic acid.............................................................................5.0mg

p-Aminobenzoic acid.................................................................5.0mg

Riboflavin ..................................................................................5.0mg

Thiamine·HCl ............................................................................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.

Reducing Agent Solution:

Composition per 10.0mL:

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

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

Preparation of Reducing Agent Solution: Add 10.0mL of dis

tilled/deionized water to a flask. Gently heat and bring to boiling. Con

tinue to boil for 1 min while sparging with 100% N2. Cool to room

temperature. Add L-cysteine·HCl·H2O. Mix thoroughly. Adjust pH to

9 with 5N NaOH. Add Na2S·9H2O. Mix thoroughly. Autoclave for 10

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 80%

N2 + 20% CO2. Add components, except reducing agent solution, to

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

Sparge with 80% N2 + 20% CO2. Anaerobically distribute into tubes or

flasks. Autoclave for 15 min at 15 psi pressure–121°C. Using a sy

ringe, aseptically and anaerobically add sterile reducing agent to each

tube (10.0mL per liter of medium).

Use: For the cultivation and maintenance of Sporomusa species.

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Sporomusa Medium