Marine Thermococcus Medium (DSMZ Medium 760)

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Marine Thermococcus Medium
(DSMZ Medium 760)

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..........................................................................................19.45g
MgCl 2 ............................................................................................8.8g
Sulfur ............................................................................................5.0g
Peptone..........................................................................................5.0g
Na 2 SO 3 ........................................................................................3.24g
CaCl 2 .............................................................................................1.8g
Yeast extract..................................................................................1.0g
KCl..............................................................................................0.55g
NaHCO 3 ......................................................................................0.16g
Ferric citrate..................................................................................0.1g
KBr..............................................................................................0.08g
SrCl 2 ............................................................................................0.03g
H 3 BO 3 .........................................................................................0.02g
Na 2 HPO 4 ....................................................................................8.0mg
Na 2 SiO 3 ......................................................................................4.0mg
NaF.............................................................................................2.4mg
NH 4 NO 3 .....................................................................................1.6mg
Na 2 S·9H 2 O solution...................................................................0.5mL
pH 6.0 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................1.0g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Neutralize to pH 7.0 with sterile HCl.
Preparation of Medium: Add components, except sulfur and
Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Filter through normal filter paper. An iron sedi-
ment will collect in the filter. Gently heat while stirring and bring to
boiling. Boil for 5 min. Cool under an anaerobic gas mixture of N 2 . Ad-
just pH to 6.0. Distribute the medium into Hungate tubes or serum bot-
tles containing finely divided sulfur (0.5% w/v). Seal the tubes or
bottles under the same anaerobic gas used when cooling the medium.
Sterilize the medium at 100°C for 3 hr on 3 consecutive days. Reduce
the medium by adding 10% neutralized Na 2 S·9H 2 O solution to a final
concentration of 0.05%. The medium should not give a heavy black
precipitate; if it does the iron sediment was not adequately removed by
filtering in the initial stages and the medium should be made again,

making sure that the iron is removed by filtering.

Use: For the cultivation and maintenance of Thermococcus aegaeus
DSM 12767.