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培养基配方

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Hydroxybenzoate Medium

[所属分类:培养基配方] [发布时间:2021-11-23] [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 1002.0mL:

Solution A................................................... 920.0mL

Solution B ..................................................... 50.0mL

Solution C ....................................................... 1.0mL

Solution D....................................................... 1.0mL

Solution E ..................................................... 10.0mL

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

Solution G..................................................... 10.0mL

pH 7.2–7.5 at 25°C

Solution A:

Composition per 920.0mL:

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

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

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

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

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

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

Preparation of Solution A: Add components to

distilled/deionized water and bring volume to

920.0mL. Mix thoroughly. Sparge with 80% N2 +

20% CO2. Autoclave for 15 min at 15 psi pressure–

121°C.

Solution B:

Composition per 50.0mL:

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

Preparation of Solution B: Add NaHCO3 to dis

tilled/deionized water and bring volume to 50.0mL.

Mix thoroughly. Sparge with 80% N2 + 20% CO2.

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

Solution C (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 C (Trace Elements So

lution 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 com

ponents. Mix thoroughly. Gas under 80% N2 + 20%

CO2. Autoclave for 15 min at 15 psi pressure–121°C.

Solution D:

Composition per 10.0mL:

NaOH...............................................................5.0mg

Na2WO4·2H2O ............................................... 40.0μg

Na2SeO3·5H2O............................................... 30.0μg

Preparation of Solution D: Add components to

distilled/deionized water and bring volume to

10.0mL. Mix thoroughly. Sparge with 80% N2 + 20%

CO2. Autoclave for 15 min at 15 psi pressure–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 components to distilled/deionized water and

bring volume to 1.0L. Mix thoroughly. Sparge with

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

pressure–121°C.

Solution F:

Composition per 10.0mL:

Sodium dihydroxybenzoate ................................0.4g

Preparation of Solution F: Add sodium dihy

droxybenzoate to distilled/deionized water and bring

volume to 10.0mL. Mix thoroughly. Filter sterilize.

Sparge with 80% N2 + 20% CO2.

Solution G:

Composition per 10.0mL:

Na2S·9H2O........................................................0.36g

Preparation of Solution G: Add Na2S·9H2O to

distilled/deionized water and bring volume to

10.0mL. Mix thoroughly. Sparge with 80% N2 + 20%

CO2. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Prepare and dispense me

dium under 80% N2 + 20% CO2. Aseptically and anaer

obically combine 920.0mL of sterile solution A,

50.0mL of sterile solution B, 1.0mL of sterile solution

C, 1.0mL of sterile solution D, 10.0mL of sterile solu

tion E, 10.0mL of sterile solution F, and 10.0mL of ster

ile solution G. Mix thoroughly. Aseptically and

anaerobically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Clost

ridium species.

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Hydroxybenzoate Medium