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Thiocapsa Medium

[所属分类:培养基配方] [发布时间:2021-9-14] [发布人:网站管理员2] [阅读次数:] [返回]

Thiocapsa Medium

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

Composition per 127.0mL:

Solution 1.................................................................................76.2mL

Solution 2 + Solution 3 ............................................................44.8mL

Solution 4...................................................................................6.0mL

Solution 1:

Composition per 2.5L:

NaCl .......................................................................................... 39.68g

CaCl2............................................................................................. 2.0g

Preparation of Solution 1: Add components to distilled/deionized

water and bring volume to 2.5L. Distribute in 80.0mL volumes into

127.0mL screw-capped bottles. Autoclave for 15 min at 15 psi pres

sure–121°C.

Solution 2:

Composition per 100.0mL:

Sodium ascorbate.......................................................................... 2.4g

KC1............................................................................................... 1.0g

KH2PO4......................................................................................... 1.0g

MgCl2·6H2O.................................................................................. 0.8g

NH4Cl ........................................................................................... 0.8g

Heavy metal solution ...............................................................50.0mL

Vitamin solution.......................................................................15.0mL

Vitamin B12 solution ..................................................................3.0mL

Preparation of Solution 2: Add components to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly.

Heavy Metal Solution:

Composition per liter:

Ethylenediamine tetraacetate (EDTA).......................................... 1.5g

FeSO4·7H2O.................................................................................. 0.2g

ZnSO4·7H2O ................................................................................. 0.1g

MnCl2·4H2O ............................................................................... 0.02g

Modified Hoagland trace elements solution..............................6.0mL

Preparation of Heavy Metal Solution: Dissolve EDTA in

approximately 800.0mL of distilled/deionized water. Add remaining

components. Bring volume to 1.0L with distilled/deionized water. Mix

thoroughly.

Modified Hoagland Trace Elements Solution:

Composition per 3.6L:

H3BO3 ......................................................................................... 11.0g

MnCl2·4H2O ................................................................................. 7.0g

AlCl3 ............................................................................................. 1.0g

CoCl2 ............................................................................................ 1.0g

CuCl2 ............................................................................................ 1.0g

KI .................................................................................................. 1.0g

NiCl2 ............................................................................................. 1.0g

ZnCl2............................................................................................. 1.0g

BaCl2............................................................................................. 0.5g

KBr ............................................................................................... 0.5g

LiCl............................................................................................... 0.5g

Na2MoO4 ...................................................................................... 0.5g

SeCl4 ............................................................................................. 0.5g

SnCl2·2H2O................................................................................... 0.5g

NaVO3·H2O .................................................................................. 0.1g

Preparation of Modified Hoagland Trace Elements Solu

tion: Prepare each component as a separate solution. Dissolve each

salt in approximately 100.0mL of distilled/deionized water. Adjust the

pH of each solution to below 7.0. Combine all the salt solutions and

bring the volume to 3.6L with distilled/deionized water. Adjust the pH

to 3–4. A yellow precipitate may form after mixing. After a few days,

it will turn into a fine white precipitate. Mix the solution thoroughly be

fore using.

Vitamin Solution:

Composition per 100.0mL:

Pyridoxamine·2HCl ................................................................... 5.0mg

Nicotinic acid............................................................................. 2.0mg

Thiamine.................................................................................... 1.0mg

Pantothenic acid......................................................................... 0.5mg

Biotin ......................................................................................... 0.2mg

p-Aminobenzoic acid................................................................. 0.1mg

Preparation of Vitamin Solution: Add components to distilled/

deionized water and bring volume to 100.0mL. Mix thoroughly.

Vitamin B12 Solution:

Composition per 100.0mL:

Vitamin B12 (cyanocobalamin) .................................................. 2.0mg

Preparation of Vitamin B12 Solution: Add vitamin B12 to dis

tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.

Solution 3:

Composition per 900.0mL:

NaHCO3 ........................................................................................ 4.5g

Preparation of Solution 3: Add NaHCO3 to distilled/deionized

water and bring volume to 900.0mL. Mix thoroughly. Bubble 100%

CO2 through the solution for 30 min. After CO2 saturation of solution

3, add solution 2 and immediately filter the mixture through a Seitz fil

ter (or a Millipore) using positive CO2 pressure to push the liquid

through.

Solution 4:

Composition per 200.0mL:

Na2S·9H2O.................................................................................... 3.0g

Preparation of Solution 4: Add Na2S·9H2O to distilled/deionized

water and bring volume to 200.0mL. Add a magnetic stir bar to the

flask. Autoclave for 15 min at 15 psi pressure–121°C. On a magnetic

stirrer, slowly add 2.0mL of sterile 2M H2SO4. This partially neutraliz

es the solution. The solution should turn yellow. H2S gas will be liber

ated; neutralization and distribution of the solution should be done as

rapidly as possible under adequate ventilation.

Preparation of Medium: To the 80.0mL of sterile solution 1 in

screw-capped bottles, add combined solutions 2 and 3 immediately af

ter filtration and fill bottles to capacity. Mix thoroughly. Aseptically re

move 6.0mL of the medium from the bottles and replace it with 6.0mL

of neutralized solution 4. Let stand for 24 hr. The medium should form

a fine white precipitate before using. To inoculate, remove 6.0mL of

the completed medium from the bottles and replace it with 6.0mL of

inoculum.

Use: For the cultivation and maintenance of a variety of Thiocapsa

species.

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Thiocapsa Medium