Methanospirillum SK Medium

[所属分类:培养基配方] [发布时间:2021-5-27] [发布人:网站管理员2] [阅读次数:] [返回]

Methanospirillum SK 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:

NaCl............................................................................................18.0g
Isopropanol ...................................................................................7.5g
NaHCO 3 ........................................................................................5.0g
MgCl 2 ·6H 2 O .................................................................................4.0g
MgSO 4 ·7H 2 O............................................................................. 3.45g
Trypticase™..................................................................................2.0g
Yeast extract..................................................................................2.0g
Sodium acetate..............................................................................1.0g
L -Cysteine·HCl .............................................................................0.5g
KCl............................................................................................0.335g
NH 4 Cl .........................................................................................0.25g
CaCl 2 ·2H 2 O ................................................................................0.14g
K 2 HPO 4 .......................................................................................0.14g
Fe(NH 4 ) 2 (SO 4 ) 2 ·7H 2 O...............................................................2.0mg
Resazurin...................................................................................1.0mg
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 7.0–7.4 at 25°C
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 approximately 500.0mL of distilled/deionized water. Dissolve
by adding KOH and adjust pH to 6.5. Add remaining components.
Bring volume to 1.0L with additional distilled/deionized water. Adjust
pH to 7.0 with KOH.
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. Sparge
with 80% N 2 + 20% CO 2 .
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.4g
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 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Preparation of Medium: Add components, except Na 2 S·9H 2 O so-
lution, to distilled/deionized water and bring volume to 990.0mL. Mix
thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at
15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL of
sterile Na 2 S·9H 2 O solution. Mix thoroughly.
Use: For the cultivation and maintenance of Methanospirillum hun-
gatei.