Methanobacterium 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 Medium

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

Composition per 1010.0mL:

NaHCO 3 ........................................................................................4.0g
Sodium formate.............................................................................2.0g
Sodium acetate..............................................................................1.0g
Yeast extract..................................................................................1.0g
L -Cysteine·HCl..............................................................................0.5g
KH 2 PO 4 .........................................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.5g
MgSO 4 ·7H 2 O............................................................................... 0.4g
NaCl..............................................................................................0.4g
NH 4 Cl ...........................................................................................0.4g
CaCl 2 ·2H 2 O.................................................................................0.05g
FeSO 4 ·7H 2 O...............................................................................2.0mg
Resazurin ...................................................................................1.0mg
Sludge fluid..............................................................................50.0mL
Fatty acid mixture....................................................................20.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 6.7 ± 0.2 at 25°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.
Fatty Acid Mixture:
Composition per 20.0mL:
α-Methylbutyric acid....................................................................0.5g
Isobutyric acid ..............................................................................0.5g
Isovaleric acid...............................................................................0.5g
Valeric acid ...................................................................................0.5g
Preparation of Fatty Acid Mixture : Add components to distilled/
deionized water and bring volume to 20.0mL. Mix thoroughly. Adjust
pH to 7.5 with concentrated NaOH.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O.......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium anaerobi-
cally under 80% H 2 + 20% CO 2 . Add components to distilled/deion-
ized water and bring volume to 1010.0mL. Mix thoroughly. Sparge
with 80% H 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Use: For the cultivation and maintenance of Methanobacterium spe-
cies.