编号：
批号：

培养基配方

您现在的位置：网站首页 >> 培养基配方

VEN CHI2 Medium (DSMZ Medium 293a)

[所属分类:培养基配方] [发布时间:2021-10-9] [发布人:网站管理员2] [阅读次数:] [返回]

VEN CHI2 Medium

(DSMZ Medium 293a)

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 ............................................................................................ 20.0g

MgCl2·6H2O.................................................................................. 3.0g

KCl................................................................................................ 0.5g

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

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

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

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

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

Na2-succinate solution .............................................................10.0mL

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

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

Quinic acid solution .................................................................10.0mL

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

pH 7.2 ± 0.2 at 25°C

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................. 0.36g

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

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

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

to room temperature.

NaHCO3 Solution:

Composition per 10.0mL:

NaHCO3 ........................................................................................ 2.5g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

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

with 80% N2 + 20% CO2. Filter sterilize.

Na2-succinate Solution:

Composition per 10.0mL:

Na2-succinate .............................................................................. 3.25g

Preparation of Na2-succinate Solution: Add Na2-succinate to

distilled/deionized water and bring volume to 10.0mL. Mix thorough

ly. Sparge with 100% N2. 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 80% H2 + 20% CO2. Filter sterilize.

Quinic Acid Solution:

Composition per 10.0mL:

Quinic acid.................................................................................... 1.0g

Preparation of Quinic Acid Solution: Add quinic acid to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Adjust pH to 7.0. Sparge with 80% N2 + 20% CO2. 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. Sparge with 80% N2 + 20% CO2. Filter sterilize.

Preparation of Medium: Prepare and dispense medium under 80%

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

tion, Na2-succinate solution, Na2S·9H2O solution, vitamin solution,

quinic acid solution, and trace elements solution SL-10, to distilled/de

ionized water and bring volume to 949.0mL. Mix thoroughly. Adjust pH

to 7.2. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min at 15 psi

pressure–121°C. Aseptically and anaerobically add 10.0mL NaHCO3

solution, 10.0mL Na2-succinate solution, 10.0mL Na2S·9H2O solution,

10.0mL vitamin solution, 10.0mL quinic acid solution, and 1.0mL

trace elements solution SL-10. Mix thoroughly. Aseptically and anaer

obically distribute into sterile tubes or bottles. After inoculation, flush

and repressurize the gas head space of culture bottles with sterile 80%

N2 + 20% CO2 to 1 bar overpressure.

Use: For the cultivation of Ilyobacter insuetus.

在线客服

VEN CHI2 Medium (DSMZ Medium 293a)