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Sporomusa Medium, Modified

[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]

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

Composition per liter:

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

Fructose solution......................................................................50.0mL

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

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

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

pH 7.0 ± 0.2 at 25°C

Fructose Solution:

Composition per 50.0mL:

Fructose....................................................................................... 10.0g

Preparation of Fructose Solution: Add fructose to distilled/de

ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge

under 100% N2 gas for 3 min. Filter sterilize. Store under N2 gas.

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. Boil under N2 gas for 1 min. Cool to

room temperature. Add L-cysteine·HCl·H2O and dissolve. Adjust to

pH 9 with 5N NaOH. Add washed Na2S·9H2O and dissolve. Mix thor

oughly. 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: Add components, except NaHCO3 and

reducing agent solution, to distilled/deionized water and bring volume

to 940.0mL. Gently heat and bring to boiling. Continue boiling for 3

min. Cool to room temperature under 80% N2 + 20% CO2. Add solid

NaHCO3 and bring pH to 7.0 by gassing. Distribute anaerobically un

der 80% N2 + 20% CO2 into tubes or flasks. Autoclave for 15 min at

15 psi pressure–121°C. Prior to inoculation of cultures, aseptically and

anaerobically add 0.1mL of sterile reducing agent solution and 0.5mL

of sterile fructose solution to each tube containing 9.4mL of sterile bas

al medium.

Use: For the cultivation and maintenance of Sporomusa acidovorans.

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Sporomusa Medium, Modified