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Syntrophomonas bryantii Medium

[所属分类:培养基配方] [发布时间:2021-9-1] [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 1026.0mL:

Solution A..............................................................................916.0mL

Solution B ................................................................................70.0mL

Solution C ................................................................................10.0mL

Solution D ................................................................................10.0mL

Sodium laurate solution ...........................................................10.0mL

CaCl2·2H2O solution................................................................10.0mL

pH 7.2 ± 0.2 at 25°C

Solution A:

Composition per 916.0mL:

PIPES (piperazine-N,N′-bis

[2-ethanesulfonic acid]) buffer ........................................... 15.12g

Na2SO4.......................................................................................... 2.8g

Butyric acid................................................................................... 1.7g

Pancreatic digest of casein............................................................ 1.0g

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

Mineral solution.......................................................................50.0mL

Rumen fluid, clarified ..............................................................50.0mL

Vitamin solution.........................................................................5.0mL

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

Mineral Solution:

Composition per liter:

Nitrilotriacetic acid ..................................................................... 12.5g

NaCl .............................................................................................. 1.0g

FeCl3·4H2O................................................................................... 0.2g

MnCl2·4H2O ................................................................................. 0.1g

CaCl2·2H2O .................................................................................. 0.1g

ZnCl2............................................................................................. 0.1g

CuCl2 .......................................................................................... 0.02g

Na2SeO3 ...................................................................................... 0.02g

CoCl2·6H2O .............................................................................. 0.017g

H3BO3......................................................................................... 0.01g

Na2MoO4·2H2O.......................................................................... 0.01g

Preparation of Mineral Solution: Add nitrilotriacetic acid to

500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add

remaining components. Add distilled/deionized water to 1.0L. Mix

thoroughly.

Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ..........................................................................6.2mg

Nicotinic acid.............................................................................2.5mg

Thiamine·HCl ..........................................................................1.25mg

p-Aminobenzoic acid...............................................................1.25mg

Pantothenic acid.......................................................................0.62mg

Biotin .......................................................................................0.25mg

Preparation of Vitamin Solution: Add components to distilled/

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

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 Solution A: Add components to distilled/deionized

water and bring volume to 916.0mL. Adjust pH to 7.2. Gently heat and

bring to boiling. Continue boiling for a few minutes. Allow to cool to

room temperature under 80% N2 + 20% CO2. Distribute into bottles

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

121°C.

Solution B:

Composition per 70.0mL:

NaHCO3........................................................................................ 3.5g

Preparation of Solution B: Add NaHCO3 to distilled/deionized

water and bring volume to 70.0mL. Mix thoroughly. Filter sterilize.

Sparge with 80% N2 + 20% CO2 for 15 min.

Solution C:

Composition per 10.0mL:

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

Preparation of Solution C: Add L-cysteine·HCl to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with

100% N2 for 3–4 min. Autoclave under 100% N2 for 15 min at 15 psi

pressure–121°C.

Solution D:

Composition per 10.0mL:

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

Preparation of Solution D: Add Na2S·9H2O to distilled/deionized

water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%

N2 for 3–4 min. Autoclave under 100% N2 for 15 min at 15 psi pres

sure–121°C.

Sodium Laurate Solution:

Composition per 10.0mL:

Sodium laurate............................................................................ 2.78g

Preparation of Sodium Laurate Solution: Add sodium laurate

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

oughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C.

CaCl2·2H2O Solution:

Composition per 40.0mL:

CaCl2·2H2O ................................................................................ 1.84g

Preparation of CaCl2·2H2O Solution: Add CaCl2·2H2O to dis

tilled/deionized water and bring volume to 40.0mL. Mix thoroughly.

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

Preparation of Medium: To 916.0mL of sterile solution A, add

70.0mL of sterile solution B, 10.0mL of sterile solution C, and 10.0mL

of sterile solution D. Mix thoroughly. Prior to inoculation, aseptically

add 10.0mL of sterile sodium laurate solution and 10.0mL of sterile

CaCl2·2H2O solution. Mix thoroughly.

Use: For the cultivation and maintenance of Syntrophomonas sapo

vorans.

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Syntrophomonas bryantii Medium