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TMBS4 Medium

[所屬分類:培養(yǎng)基配方] [發(fā)布時間:2021-9-16] [發(fā)布人:網(wǎng)站管理員2] [閱讀次數(shù):] [返回]

TMBS4 Medium

山東拓普生物工程有限公司培養(yǎng)基配方 http://qp8008.cn

Composition per 1002.0mL:

Solution A ..............................................................................870.0mL

Solution C ..............................................................................100.0mL

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

Solution E (Vitamin solution)..................................................10.0mL

Solution F.................................................................................10.0mL

Solution B (Trace elements solution SL-10) .............................1.0mL

Solution G..................................................................................1.0mL

pH 7.1–7.4 at 25°C

Solution A:

Composition per 870.0mL:

Na2SO4.......................................................................................... 3.0g

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

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

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

NH4Cl ........................................................................................... 0.3g

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

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

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

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 870.0mL. Mix thoroughly. Gently heat and

bring to boiling. Continue boiling for 3–4 min. Allow to cool to room

temperature while gassing under 80% N2 + 20% CO2. Continue gas

sing until pH reaches below 6.0. Seal the flask under 80% N2 + 20%

CO2. Autoclave for 15 min at 15 psi pressure–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):

Add FeCl2·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis

tilled/deionized water and bring volume to 1.0L. Add remaining com

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

15 psi pressure–121°C.

Solution C:

Composition per 100.0mL:

NaHCO3........................................................................................ 5.0g

Preparation of Solution C: Add NaHCO3 to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Gas under 80% N2 + 20% CO2.

Solution D:

Composition per 10.0mL:

Syringic acid ................................................................................. 0.5g

Preparation of Solution D: Add syringic acid to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to

8 with NaOH. Gas under 100% N2. Autoclave for 15 min at 15 psi pres

sure–121°C.

Solution E (Vitamin Solution):

Composition per liter:

Pyridoxine·HCl ........................................................................ 10.0mg

Calcium DL-pantothenate........................................................... 5.0mg

Lipoic acid ................................................................................. 5.0mg

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

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

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

Thiamine·HCl ............................................................................5.0mg

Biotin .........................................................................................2.0mg

Folic acid....................................................................................2.0mg

Vitamin B12 ................................................................................0.1mg

Preparation of Solution E (Vitamin Solution): Add compo

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

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

121°C.

Solution F:

Composition per 10.0mL:

Na2S·9H2O.................................................................................. 0.25g

Na2S2O4 ...................................................................................... 0.25g

Preparation of Solution F: Add components 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.

Solution G:

Composition per liter:

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

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

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

Preparation of Solution G: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Gas under 100% N2.

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

Preparation of Medium: Aseptically and anaerobically combine

solution A with solution B, solution C, solution D, solution E, solution

F, and solution G, in that order. Mix thoroughly. Anaerobically distrib

ute into sterile tubes or flasks under 80% N2 + 20% CO2.

Use: For the cultivation and maintenance of Pelobacter species.

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TMBS4 Medium