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YPS Medium (DSMZ Medium 990)

[所屬分類:培養基配方] [發布時間:2021-10-25] [發布人:網站管理員2] [閱讀次數:] [返回]
YPS Medium
(DSMZ Medium 990)

山東拓普生物工程有限公司 培養基配方 //ljxinglong.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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