Methanococcus jannaschii Medium

[所属分类:培养基配方] [发布时间:2021-5-24] [发布人:网站管理员2] [阅读次数:] [返回]

Methanococcus jannaschii Medium

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

Composition per 1020.0mL:

PIPES (piperazine-N,N′-
bis[2-ethanesulfonic acid]) buffer ......................................15.12g
MgCl 2 ·6H 2 O..................................................................................4.3g
MgSO 4 ·7H 2 O................................................................................3.4g
NaCl..............................................................................................3.0g
NH 4 Cl .........................................................................................0.25g
K 2 HPO 4 .......................................................................................0.14g
CaCl 2 ·2H 2 O ................................................................................0.14g
KCl..............................................................................................0.33g
Minerals solution.....................................................................10.0mL
Na 2 S 2 O 3  solution......................................................................10.0mL
β-Mercaptoethanol solution.....................................................10.0mL
SeO 2  solution .............................................................................1.0mL
pH 7.0 ± 0.2 at 25°C
Minerals Solution:
Composition per liter:
Nitrilotriacetic acid.......................................................................4.5g
FeCl 2 ·4H 2 O...................................................................................0.4g
CoCl 2 ·2H 2 O................................................................................0.17g
MnCl 2 ·4H 2 O .................................................................................0.1g
ZnCl 2 .............................................................................................0.1g
NaMoO 4 ·6H 2 O.........................................................................36.0mg
CaCl 2 ·H 2 O ...............................................................................27.0mg
Preparation of Minerals Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Na 2 S 2 O 3 Solution:
Composition per 10.0mL:
Na 2 S 2 O 3 ·5H 2 O............................................................................0.63g
Preparation of Na 2 S 2 O 3 Solution:  Add Na 2 S 2 O 3 ·5H 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.
β-Mercaptoethanol Solution:
Composition per 10.0mL:
β-Mercaptoethanol......................................................................0.39g
Preparation of β-Mercaptoethanol Solution:  Add β-mercapto-
ethanol to distilled/deionized water and bring volume to 10.0mL. Mix
thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
SeO 2 Solution:
Composition per 100.0mL:
SeO 2 .......................................................................................... 0.011g
Preparation of SeO 2 Solution:  Add SeO 2 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under
100% N 2 . Add components, except Na 2 S 2 O 3 solution and β-mercapto-
ethanol solution, to distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling. Continue boiling for
3 min. Cool to room temperature while sparging with 100% N 2 . Adjust
pH to 7.0 with KOH. Anaerobically distribute 100.0mL volumes into an-
aerobic bottles. Autoclave for 15 min at 15 psi pressure–121°C. Aseptical-
ly and anaerobically add 1.0mL of sterile Na 2 S 2 O 3 solution and 1.0mL of
sterile β-mercaptoethanol solution to each bottle. Mix thoroughly. Prior to
inoculation, flush each bottle with 80% H 2 + 20% CO 2 .
Use:  For the cultivation of Methanococcus jannaschii.