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YPS Medium (DSMZ Medium 990)

[所属分类:培养基配方] [发布时间:2021-10-25] [发布人:网站管理员2] [阅读次数:] [返回]

YPS Medium

(DSMZ Medium 990)

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

Composition per liter:

Sea salts, Sigma .......................................................................... 35.0g

Sulfur, elemental ........................................................................... 5.0g

PIPES (piperazine-N,N'-bis[2-ethane-sulfonic acid]) ................ 3.46g

NH4Cl .......................................................................................... 0.5g

KH2PO4 ...................................................................................... 0.35g

CaCl2·2H2O................................................................................... 0.2g

FeCl3·6H2O ................................................................................6.7mg

Na2WO4 ....................................................................................2.9mg

Resazurin ..................................................................................0.1mg

Yeast extract solution...............................................................10.0mL

Peptone solution.......................................................................10.0mL

Sulfide solution ..........................................................................5.0mL

pH 6.8 ± 0.2 at 25°C

Sulfide Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.5g

Preparation of Sulfide Solution: Add Na2S·9H2O to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave

under 100% N2 for 15 min at 15 psi pressure–121°C. Cool to room

temperature. Adjust pH to 7.0.

Yeast Extract Solution:

Composition per 10.0mL:

Yeast extract ................................................................................. 1.0g

Preparation of Yeast Extract Solution: Add yeast extract to dis

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

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

ature. Sparge with 100% N2.

Peptone Solution:

Composition per 10.0mL:

Peptone ......................................................................................... 4.0g

Preparation of Peptone Solution: Add peptone to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for

15 min at 15 psi pressure–121°C. Cool to room temperature. Sparge

with 100% N2.

Preparation of Medium: Add components, except yeast extract,

peptone, and sulfide solutions, to distilled/deionized water and bring

volume to 975.0mL. Mix thoroughly. Gently heat and bring to boiling.

Cool to room temperature while sparging with 100% N2. Adjust pH to

6.8. Prepare the medium without the yeast extract, peptone, and sodium

sulfide. Boil the medium and cool under nitrogen. Adjust the pH to 6.8.

Dispense into Hungate tubes or serum bottles under a nitrogen atmo

sphere. Sterilize the medium at 100°C for 3 hr on 3 consecutive days.

Aseptically add the peptone and yeast extract solutions. Adding the

sterile, neutralized sulfide solution to an end concentration of 0.025%.

Final pH should be 6.8.

Use: For the cultivation of Thermococcus marinus.

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YPS Medium (DSMZ Medium 990)