Marinitoga piezophila Medium (DSMZ Medium 945)

[所属分类:培养基配方] [发布时间:2021-5-17] [发布人:网站管理员2] [阅读次数:] [返回]

Marinitoga piezophila Medium
(DSMZ Medium 945)

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............................................................................................30.0g
Sulfur..........................................................................................10.0g
Yeast extract..................................................................................5.0g
Trypticase™..................................................................................5.0g
MES............................................................................................1.95g
NH 4 Cl ...........................................................................................1.0g
Na-acetate...................................................................................0.83g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
MgCl 2 ·6H 2 O .................................................................................0.2g
CaCl 2 ·2H 2 O ..................................................................................0.1g
KCl................................................................................................0.1g
Resazurin...................................................................................0.5mg
Na 2 S·9H 2 O solution.................................................................10.0mL
Cysteine solution .....................................................................10.0mL
pH 6.0 ± 0.2 at 25°C
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O.....................................................................0.3g
Preparation of  L -Cysteine Solution:  Add  L -cysteine·HCl·H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
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. Sparge with N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
Preparation of Sulfur:  Sterilize sulfur by steaming for 3 hr on each
of 3 successive days.
Preparation of Medium: Add components, except sulfur, cysteine
solution, and Na 2 S·9H 2 O solution, to 980.0mL distilled/deionized wa-
ter. Mix thoroughly. Sparge for 30 min with 100% N 2 . Adjust pH to 6.0
with concentrated NaOH. Distribute under 100% N 2 into anaerobic
tubes or bottles containing appropriate amounts of sterile sulfur (1g
steam-sterilized sulfur per 100mL medium). Autoclave for 20 min at
110°C. Cool to room temperature. Aseptically and anaerobically add
10.0mL sterile Na 2 S·9H 2 O solution and 10.0mL sterile cysteine solu-
tion per liter medium. Mix thoroughly.
Use:  For the cultivation of Marinitoga piezophila.