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SWM Medium

[所属分类:培养基配方] [发布时间:2021-8-31] [发布人:网站管理员2] [阅读次数:] [返回]

SWM Medium

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

Composition per 1014.0mL:

Solution A ..............................................................................940.0mL

Solution E (NaHCO3 solution) ................................................50.0mL

Solution F (Substrate solution) ................................................10.0mL

Solution G (Na2S·9H2O solution)............................................10.0mL

Solution B (Trace elements solution SL-10)..............................2.0mL

Solution C (Seven vitamin solution)..........................................1.0mL

Solution D (Selenite-tungstate solution)....................................1.0mL

pH 7.2–7.4 at 25°C

Solution A:

Composition per 940.0mL:

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

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

MgCl2·6H2O.................................................................................. 0.4g

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

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

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

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

Preparation of Solution A: Prepare and dispense under 80% N2 +

20% CO2. Add components to distilled/deionized water and bring vol

ume to 940.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pres

sure–121°C.

Solution B (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 Solution B (Trace Elements Solution SL-10):

Prepare and dispense under 100% N2. 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 thoroughly. Auto

clave for 15 min at 15 psi pressure–121°C.

Solution C (Seven Vitamin Solution):

Composition per liter:

Pyridoxine·HCl ......................................................................300.0mg

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

Thiamine·HCl ........................................................................200.0mg

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

Cyanocobalamine...................................................................100.0mg

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

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

Preparation of Solution C (Seven Vitamin Solution): Add

components to distilled/deionized water and bring volume to 1.0L. Mix

thoroughly. Filter sterilize. Sparge with 100% N2.

Solution D (Selenite-Tungstate Solution):

Composition per liter:

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

Na2WO4·2H2O........................................................................... 4.0mg

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

Preparation of Solution D (Selenite-Tungstate Solution):

Add components to distilled/deionized water and bring volume to

1.0L. Mix thoroughly. Filter sterilize. Sparge with 100% N2.

Solution E (NaHCO3 Solution):

Composition per 50.0mL:

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

Preparation of Solution E (NaHCO3 Solution): Add NaHCO3

to distilled/deionized water and bring volume to 50.0mL. Mix thor

oughly. Gas under 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C.

Solution F (Substrate Solution):

Composition per 10.0mL:

Pyrogallol...................................................................................... 0.5g

Preparation of Solution F (Substrate Solution): Add pyrogallol

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

oughly. Gas under 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C.

Solution G (Na2S·9H2O Solution):

Composition per 10.0mL:

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

Preparation of Solution G (Na2S·9H2O Solution): Add

Na2S·9H2O to distilled/deionized water and bring volume to 10.0mL.

Mix thoroughly. Gas under 100% N2. Autoclave for 15 min at 15 psi

pressure–121°C.

Preparation of Medium: To 940.0mL of sterile solution A, asepti

cally and anaerobically add 1.0mL of sterile solution B, 1.0mL of ster

ile solution C, 1.0mL of sterile solution D, 50.0mL of sterile solution

E, 10.0mL of sterile solution F, and 10.0mL of sterile solution G. Mix

thoroughly. Aseptically and anaerobically distribute into sterile tubes

or flasks.

Use: For the cultivation and maintenance of Pelobacter massiliensis.

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SWM Medium