Methanobacterium Mass-Culturing Medium

[所属分类:培养基配方] [发布时间:2021-5-21] [发布人:网站管理员2] [阅读次数:] [返回]

Methanobacterium Mass-Culturing Medium

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

Composition per liter:

NH 4 Cl ...........................................................................................2.0g
NaCl..............................................................................................0.6g
KH 2 PO 4 .......................................................................................0.42g
K 2 HPO 4 .......................................................................................0.23g
Na 2 CO 3 .......................................................................................0.16g
MgCl 2 ·2H 2 O ...............................................................................0.04g
CaCl 2 ·2H 2 O ................................................................................0.03g
Resazurin..................................................................................0.001g
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution...........................................................10.0mL
Na 2 S·9H 2 O Solution:
Composition per 100.0mL:
NaOH....................................................................................... 1 pellet
Na 2 S·9H 2 O....................................................................................2.5g
Preparation of Na 2 S·9H 2 O Solution:  Bring 100.0mL of dis-
tilled/deionized water to boiling. Cool to room temperature while
sparging with 100%N 2 . Dissolve 1 pellet of NaOH in the anaerobic wa-
ter. Weigh out a little more than 2.5g of Na 2 S·9H 2 O. Briefly rinse the
crystals in distilled/deionized water. Dry the crystals by blotting on pa-
per towels or filter paper. Add 2.5g of washed Na 2 S·9H 2 O crystals to
100.0mL of anaerobic NaOH solution. Distribute into serum bottles fit-
ted with butyl rubber stoppers and aluminum seals. Do not grease stop-
pers. Pressurize to 60kPa with 100% N 2 . Autoclave for 15 min at 15 psi
pressure–121°C. Store at room temperature in an anaerobic chamber.
Trace Elements Solution:
Composition per liter:
Sodium nitrilotriacetate...............................................................1.67g
CoCl 2 ·6H 2 O ................................................................................0.18g
FeCl 2 ·4H 2 O.................................................................................0.14g
MnCl 2 ·4H 2 O................................................................................0.09g
NiCl 2 ·6H 2 O.................................................................................0.09g
ZnCl 2 ...........................................................................................0.09g
CaCl 2 ...........................................................................................0.06g
Na 2 MoO 4 ·2H 2 O ........................................................................0.046g
CuSO 4 .......................................................................................0.045g
Preparation of Trace Elements Solution:  Add sodium nitrilotri-
acetate to 500.0mL of distilled/deionized water. Adjust pH to 6.5. Add
remaining components. Add distilled/deionized water to 1.0L. Adjust
pH to 7.0.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 . Add components, except Na 2 CO 3 and Na 2 S·9H 2 O so-
lution, to distilled/deionized water and bring volume to 990.0mL. Mix
thoroughly. Gently heat and bring to boiling. Continue boiling for 3
min. Cool to room temperature while sparging with 80% H 2 + 20%
CO 2 . Add Na 2 CO 3 . Mix thoroughly. Anaerobically distribute 9.9mL
volumes into anaerobic tubes. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 0.1mL of sterile Na 2 S·9H 2 O
solution to each tube. Mix thoroughly.
Use:  For the cultivation of Methanobacterium species.