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Tissierella creatinophila Medium (DSMZ Medium 824)

[所属分类:培养基配方] [发布时间:2021-9-16] [发布人:网站管理员2] [阅读次数:] [返回]

Tissierella creatinophila Medium

(DSMZ Medium 824)

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

Composition per 1022.0mL:

Creatine......................................................................................... 3.8g

Na-formate .................................................................................. 2.72g

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

KCl................................................................................................ 1.0g

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

NaCl .............................................................................................. 0.5g

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

KH2PO4......................................................................................... 0.2g

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

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

Na2SeO3·5H2O.........................................................................0.26mg

NaHCO3 solution .....................................................................40.0mL

Na2S·9H2O solution .................................................................20.0mL

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

Seven vitamin solution...............................................................1.0mL

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

pH 7.5–7.8 ± 0.2 at 25°C

Na2S·9H2O Solution:

Composition per 20.0mL:

Na2S·9H2O.................................................................................... 0.6g

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

tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.

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

to room temperature.

NaHCO3 Solution:

Composition per 100.0mL:

NaHCO3 ...................................................................................... 10.0g

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.

Vitamin Solution:

Composition per liter:

Pyridoxine-HCl........................................................................10.0mg

Thiamine-HCl·2H2O..................................................................5.0mg

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

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

D-Ca-pantothenate......................................................................5.0mg

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

Lipoic acid .................................................................................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. Sparge

with 100% N2. Filter sterilize.

Seven Vitamin Solution:

Composition per liter:

Pyridoxine hydrochloride ......................................................300.0mg

Thiamine-HCl·2H2O..............................................................200.0mg

Nicotinic acid.........................................................................200.0mg

Vitamin B12 ............................................................................100.0mg

Calcium pantothenate ............................................................100.0mg

p-Aminobenzoic acid...............................................................80.0mg

D(+)-Biotin...............................................................................20.0mg

Preparation of Seven Vitamin Solution: Add components to dis

tilled/deionized water and bring volume to 1.0L. Sparge with 100% N2.

Mix thoroughly. Filter sterilize.

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 NaHCO3 solution, Na2S·9H2O solution,

seven vitamin solution, and vitamin solution, to 950.0mL distilled/de

ionized water. Mix thoroughly. Sparge with 100% N2 + 20% CO2. Adjust

pH to 7.6. Dispense 10.0mL aliquots into bottles. Autoclave for 15 min

at 15 psi pressure–121°C. For each 10.0mL medium aseptically and an

aerobically inject from sterile stock solutions 0.4mL NaHCO3 solution,

0.2mL Na2S·9H2O solution, 0.1mL vitamin solution, and 0.01mL sev

en vitamin solution. Final pH should be 7.5–7.8.

Use: For the cultivation of Tissierella creatinophila.

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Tissierella creatinophila Medium (DSMZ Medium 824)