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Sulfate-Reducing Bacteria Enrichment Medium

[所属分类:培养基配方] [发布时间:2021-8-30] [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 1018.0mL:

Solution 1...............................................................................970.0mL

Solution 4.................................................................................30.0mL

Solution 6A, 6B, 6C, 6D, or 6E...............................................10.0mL

Solution 5...................................................................................3.0mL

Solution 2...................................................................................1.0mL

Solution 3...................................................................................1.0mL

Solution 7...................................................................................1.0mL

Solution 8...................................................................................1.0mL

Solution 9...................................................................................1.0mL

pH 7.2 ± 0.2 at 25°C

Solution 1:

Composition per 970.0mL:

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

NaCl.............................................................................................. 1.2g

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

KCl................................................................................................ 0.3g

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

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

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

Preparation of Solution 1: Add components to distilled/deionized

water and bring volume to 970.0mL. Mix thoroughly. Autoclave for 30

min at 15 psi pressure–121°C. Cool to 25°C under 90% N2 + 10% CO2.

Solution 2:

Composition per liter:

FeCl2·4H2O ................................................................................... 1.5g

CoCl2·6H2O ................................................................................ 0.12g

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

ZnCl2........................................................................................... 0.07g

H3BO3 ......................................................................................... 0.06g

Na2MoO4·2H2O ........................................................................ 0.025g

NiCl2·6H2O ............................................................................... 0.025g

CuCl2·2H2O .............................................................................. 0.015g

HCl (25% solution)....................................................................6.5mL

Preparation of Solution 2: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min

at 15 psi pressure–121°C. Cool to 25°C.

Solution 3:

Composition per liter:

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

Na2SeO3 .....................................................................................3.0mg

Preparation of Solution 3: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min

at 15 psi pressure–121°C. Cool to 25°C.

Solution 4:

Composition per 100.0mL:

NaHCO3 ........................................................................................ 8.5g

Preparation of Solution 4: Add NaHCO3 to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Saturate with

100% CO2. Filter sterilize. Aseptically add solution to sterile, gas

tight, screw-capped bottles.

Solution 5:

Composition per 100.0mL:

Na2S·9H2O.................................................................................. 12.0g

Preparation of Solution 5: Add Na2S·9H2O to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Add solution to

gas-tight, screw-capped bottles. Gas under 100% N2 for 20 min. Close

caps tightly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

25°C.

Solution 6A:

Composition per 100.0mL:

Sodium acetate·3H2O.................................................................. 20.0g

Preparation of Solution 6A: Add sodium acetate·3H2O to dis

tilled/deionized water and bring volume to 100.0mL. Autoclave for 15

min at 15 psi pressure–121°C. Cool to 25°C.

Solution 6B:

Composition per 100.0mL:

n-Butyric acid ............................................................................... 8.0g

Preparation of Solution 6B: Add n-butyric acid to distilled/deion

ized water and bring volume to 100.0mL. Adjust pH to 9.0 with NaOH.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Solution 6C:

Composition per 100.0mL:

Propionic acid ............................................................................... 7.0g

Preparation of Solution 6C: Add propionic acid to 100.0mL of

distilled/deionized water. Adjust pH to 9.0 with NaOH. Autoclave for

15 min at 15 psi pressure–121°C. Cool to 25°C.

Solution 6D:

Composition per 100.0mL:

Benzoic acid.................................................................................. 5.0g

Preparation of Solution 6D: Add benzoic acid to distilled/deion

ized water and bring volume to 100.0mL. Adjust pH to 9.0 with NaOH.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Solution 6E:

Composition per 100.0mL:

n-Palmitic acid.............................................................................. 5.0g

NaOH.......................................................................................... 0.78g

Preparation of Solution 6E: Add n-palmitic acid and NaOH to

distilled/deionized water and bring volume to 100.0mL. Heat in a water

bath until clear. Autoclave for 15 min at 15 psi pressure–121°C. Cool

to 25°C.

Solution 7:

Composition per 100.0mL:

Thiamine..................................................................................... 0.01g

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

Vitamin B12 ................................................................................ 5.0mg

Biotin ......................................................................................... 1.0mg

Preparation of Solution 7: Add components to distilled/deionized

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

Solution 8:

Composition per liter:

Succinic acid................................................................................. 0.6g

Isobutyric acid .............................................................................. 0.5g

2-Methylbutyric acid .................................................................... 0.5g

3-Methylbutyric acid .................................................................... 0.5g

Valeric acid ................................................................................... 0.5g

Caproic acid.................................................................................. 0.2g

Preparation of Solution 8: Add components to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Adjust pH to 9.0

with NaOH. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

25°C.

Solution 9:

Composition per 100.0mL:

Na2S2O4 ........................................................................................ 3.0g

Preparation of Solution 9: Add Na2S2O4 to 100.0mL of O2-free

distilled/deionized water. Mix thoroughly. Anaerobically filter steril

ize.

Preparation Medium: To 970.0mL of cooled, sterile solution 1,

aseptically and anaerobically add 1.0mL of sterile solution 2, 1.0mL of

sterile solution 3, 30.0mL of sterile solution 4, and 3.0mL of sterile so

lution 5. Mix thoroughly. Adjust pH to 7.2 with sterile HCl solution or

sterile Na2CO3 solution. Aseptically and anaerobically distribute into

sterile screw-capped bottles in 100.0mL volumes. Add 1.0mL of solu

tion 6A, 6B, 6C, 6D, or 6E to each bottle containing 100.0mL of basal

medium. Add 0.1mL of solution 7, 0.1mL of solution 8, and 0.1mL of

solution 9 to each bottle containing 100.0mL of basal medium. Mix

thoroughly.

Use: For the isolation, cultivation, and enrichment of sulfate-reducing

bacteria.

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Sulfate-Reducing Bacteria Enrichment Medium