Methanomicrobium paynteri Medium

[所属分类:培养基配方] [发布时间:2021-5-26] [发布人:网站管理员2] [阅读次数:] [返回]

Methanomicrobium paynteri 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:

3-(N-morpholino) propane
sulfonic acid (MOPS buffer)..............................................20.93g
NaCl............................................................................................6.31g
NaHCO 3 ........................................................................................5.0g
Sodium acetate·3H 2 O..................................................................4.14g
MgSO 4 ·7H 2 O..............................................................................3.40g
MgCl 2 ·2H 2 O ...............................................................................2.75g
NH 4 Cl ...........................................................................................1.5g
KCl..............................................................................................0.34g
CaCl 2 ·2H 2 O ................................................................................0.14g
K 2 HPO 4 .......................................................................................0.14g
Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O...............................................................2.0mg
Resazurin ...................................................................................1.0mg
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
L -Cysteine·HCl solution ..........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O............................................................................... 3.0g
NaCl..............................................................................................1.0g
Nitrilotriacetic acid.......................................................................1.5g
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
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
NiCl 2 ·6H 2 O...............................................................................0.025g
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.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution : Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize. Sparge with 80% H 2 + 20% CO 2 .
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl..............................................................................0.5g
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....................................................................................0.5g
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.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 . Add components, except MOPS buffer,  L -cysteine·HCl
solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and
bring volume to 890.0mL. Mix thoroughly. Sparge with O 2 -free 80%
H 2 + 20% CO 2 . In a separate flask, add MOPS buffer to distilled/deion-
ized water and bring volume to 90.0mL. Adjust pH to 7.0 with 2M
KOH. Add the 90.0mL of MOPS solution to the 890.0mL of medium
and continue sparging with O 2 -free 80% H 2 + 20% CO 2 . Anaerobically
distribute into tubes or flasks fitted with butyl rubber stoppers. Auto-
clave for 15 min at 15 psi pressure–121°C. Anaerobically add 10.0mL
of sterile  L -cysteine·HCl solution and 10.0mL of sterile Na 2 S·9H 2 O so-
lution to each liter of medium or, using a syringe, inject the appropriate
amount of sterile  L -cysteine·HCl solution and sterile Na 2 S·9H 2 O solu-
tion into individual tubes containing medium.
Use: For the cultivation and maintenance of Methanomicrobium paynteri.