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SF1 Medium

[所属分类:培养基配方] [发布时间:2021-8-16] [发布人:网站管理员2] [阅读次数:] [返回]

SF1 Medium

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

Composition per liter:

NaCl.......................................................................................... 120.0g

MgCl2·6H2O ................................................................................. 7.0g

MgSO4·7H2O................................................................................ 6.0g

KCl................................................................................................ 3.8g

Pancreatic digest of casein............................................................ 2.0g

Yeast extract.................................................................................. 2.0g

NH4Cl ........................................................................................... 1.0g

CaCl2·2H2O .................................................................................. 0.5g

L-Cysteine·HCl ............................................................................. 0.5g

K2HPO4·3H2O ............................................................................. .0.4g

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

Na2SeO3·5H2O..........................................................................75.0μg

Na2CO3 solution.....................................................................20.0mL

Trimethylamine·HCl solution ..................................................20.0mL

Na2S·9H2O solution .................................................................10.0mL

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

NaOH (10M solution) ................................................................0.6mL

pH 7.3 ± 0.2 at 25°C

Na2CO3 Solution:

Composition per 20.0mL:

Na2CO3 ....................................................................................10.0mg

Preparation of Na2CO3 Solution: Add Na2CO3 to distilled/deion

ized water and bring volume to 20.0mL. Mix thoroughly. Sparge under

100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Store under

N2.

Trimethylamine·HCl Solution:

Composition per 20.0mL:

Trimethylamine·HCl ................................................................10.0mg

Preparation of Trimethylamine·HCl Solution: Add trimethyl

amine·HCl to distilled/deionized water and bring volume to 20.0mL.

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

pressure–121°C. Store under N2.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O...............................................................................10.0mg

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

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

121°C. Store under N2.

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

psi pressure–121°C.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2. Add components, except L-cysteine·HCl, NaOH,

Na2CO3 solution, trimethylamine·HCl solution, and Na2S·9H2O solu

tion, to distilled/deionized water and bring volume to 950.0mL. Mix

thoroughly. Gently heat and bring to boiling. Continue boiling for 5

min. Cool to room temperature while sparging with 80% N2 + 20%

CO2. Add L-cysteine·HCl and NaOH while contiuning to sparge with

80% N2 + 20% CO2. Adjust pH to 6.7. Anaerobically distribute into

tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Asep

tically and anaerobically add 20.0mL of sterile trimethylamine·HCl so

lution, 20.0mL of sterile Na2CO3 solution, and 10.0mL of sterile

Na2S·9H2O solution per 950.0mL of medium. Check that final pH is

6.7.

Use: For the cultivation and maintenance of Methanohalophilus species.

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SF1 Medium