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Thermoanaerobacterium Medium (DSMZ Medium 903)

[所属分类:培养基配方] [发布时间:2021-9-6] [发布人:网站管理员2] [阅读次数:] [返回]

Thermoanaerobacterium Medium

(DSMZ Medium 903)

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

Composition per 1030.0mL:

KH2PO4......................................................................................... 0.5g

NaCl .............................................................................................. 0.4g

MgCl2·6H2O................................................................................ 0.33g

Trypticase™................................................................................ 0.25g

CaCl2·2H2O................................................................................. 0.05g

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

Sucrose solution.......................................................................50.0mL

NaHCO3 solution .....................................................................20.0mL

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

L-Cysteine solution ..................................................................10.0mL

Selenite solution.........................................................................1.0mL

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

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

pH 7.0 ± 0.2 at 25°C

Sucrose Solution:

Composition per 50.0mL:

Sucrose.......................................................................................... 5.0g

Preparation of Sucrose Solution: Add sucrose to distilled/deion

ized water and bring volume to 50.0mL. Mix thoroughly. Sparge with

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

L-Cysteine Solution:

Composition per 10.0mL:

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

Preparation of L-Cysteine Solution: Add L-cysteine·HCl·H2O to

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

ly. 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.

NaHCO3 Solution:

Composition per 20.0mL:

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

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 20.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to 25°C. Must be prepared freshly.

Selenite Solution:

Composition per liter:

NaOH............................................................................................ 0.5g

Na2SeO3·5H2O........................................................................... 3.0mg

Preparation of Selenite 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. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min at 15

psi pressure–121°C.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2. Add components, except seven vitamin solution,

NaHCO3 solution, sucrose solution, L-cysteine-HCl·H2O solution, and

Na2S·9H2O solution, to distilled/deionized water and bring volume to

940.0mL. Mix thoroughly. Adjust pH to 7.0. Distribute into anaerobe

tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Asepti

cally and anaerobically add, per liter, 10.0mL seven vitamin solution,

20.0mL NaHCO3 solution, 50.0mL sucrose solution, 10.0mL L

cysteine-HCl·H2O solution, and 10.0mL Na2S·9H2O solution. Mix

thoroughly. The final pH should be 7.0.

Use: For the cultivation of Thermoanaerobacterium polysaccharolyt

icum and Thermoanaerobacterium zeae.

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Thermoanaerobacterium Medium (DSMZ Medium 903)