Methanogenium aggregans Medium

[所属分类:培养基配方] [发布时间:2021-5-25] [发布人:网站管理员2] [阅读次数:] [返回]

Methanogenium aggregans 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:

Sodium formate ............................................................................5.0g
NH 4 Cl ...........................................................................................1.0g
Sodium acetate..............................................................................1.0g
Trypticase™..................................................................................1.0g
Yeast extract..................................................................................1.0g
K 2 HPO 4 ·3H 2 O ..............................................................................0.4g
MgCl 2 ·6H 2 O .................................................................................0.4g
Resazurin ...................................................................................1.0mg
Mineral solution.......................................................................50.0mL
Trace elements solution...........................................................10.0mL
L -Cysteine·HCl solution ..........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Na 2 CO 3  solution.......................................................................10.0mL
Sludge fluid................................................................................5.0mL
pH 6.8 ± 0.2 at 25°C
Mineral Solution:
Composition per liter:
NaCl............................................................................................12.0g
KH 2 PO 4 .........................................................................................6.0g
(NH 4 ) 2 SO 4 .....................................................................................6.0g
MgSO 4 ·7H 2 O................................................................................2.6g
CaCl 2 ·2H 2 O.................................................................................0.16g
Preparation of Mineral Solution : Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O............................................................................... 3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O............................................................................... 0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ......................................................................... 0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Adjust pH to 7.0. Add distilled/de-
ionized water to 1.0L.
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl..............................................................................0.2g
Preparation of  L -Cysteine·HCl Solution: Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.2g
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.
Na 2 CO 3 Solution:
Composition per 10.0mL:
Na 2 CO 3 .........................................................................................1.5g
Preparation of Na 2 CO 3 Solution:  Add Na 2 CO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Sludge Fluid:
Composition per 100.0mL:
Yeast extract..................................................................................0.4g
Sludge ....................................................................................100.0mL
Preparation of Sludge Fluid : To 100.0mL of sludge from an an-
aerobic digester, add 0.4g of yeast extract. Sparge with 100% N 2 for a
few minutes. Incubate at 37°C for 24 hr. Centrifuge the sludge at
13,000 × g for 15 min. Decant the clear supernatant solution. Sparge
with 100% N 2 for a few minutes. Store in screw-capped bottles at room
temperature in the dark.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 . Add components, except  L -cysteine·HCl solution,
Na 2 S·9H 2 O solution, and Na 2 CO 3 solution, to distilled/deionized water
and bring volume to 970.0mL. Mix thoroughly. Adjust pH to 6.8. An-
aerobically distribute into tubes or flasks fitted with butyl rubber stop-
pers. Autoclave for 15 min at 15 psi pressure–121°C. Anaerobically
add 10.0mL of sterile  L -cysteine·HCl solution, 10.0mL of sterile
Na 2 S·9H 2 O solution, and 10.0mL of sterile Na 2 CO 3 solution to each li-
ter of medium or, using a syringe, inject the appropriate amount of ster-
ile  L -cysteine·HCl solution, sterile Na 2 S·9H 2 O solution, and sterile
Na 2 CO 3 solution into individual tubes containing medium.
Use: For the cultivation and maintenance of Methanocorpusculum
aggregans.