Methylophaga Agar

[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]

Methylophaga Agar

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

Composition per 103.0mL:

Agar solution............................................................................50.0mL
Mineral base, 2X......................................................................50.0mL
Solution T ..................................................................................2.0mL
Vitamin B 12 solution..................................................................1.0mL
Methanol....................................................................................0.3mL
pH 7.3 ± 0.2 at 25°C
Agar Solution:
Composition per 500.0mL:
Agar ............................................................................................15.0g
Preparation of Agar Solution:  Add agar to distilled/deionized
water and bring volume to 500.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to 50°C.
Mineral Base, 2X:
Composition per 500.0mL:
NaCl............................................................................................24.0g
MgCl 2 ·6H 2 O..................................................................................3.0g
MgSO 4 ·7H 2 O................................................................................2.0g
CaCl 2 ·2H 2 O...................................................................................1.0g
KCl................................................................................................0.5g
Bis-Tris buffer (bis[2-hydroxyethyl]amino-
tris[hydroxymethyl]-methane)............................................0.5g
Wolfe’s mineral solution..........................................................10.0mL
Preparation of Mineral Base, 2X:  Add components to distilled/
deionized water and bring volume to 500.0mL. Mix thoroughly. Adjust
pH to 7.3. Autoclave for 15 min at 15 psi pressure–121°C. Cool to
50°C.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
FeSO 4 ·7H 2 O..................................................................................0.1g
CoCl 2 ·6H 2 O..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CuSO 4 ·5H 2 O...............................................................................0.01g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L.
Solution T:
Composition per 100.0mL:
NH 4 Cl .........................................................................................10.0g
Bis-Tris buffer (bis[2-hydroxyethyl]amino-
tris[hydroxymethyl]-methane) .........................................10.0g
KH 2 PO 4 .........................................................................................0.7g
Ferric ammonium citrate...............................................................0.3g
Preparation of Solution T:  Add components to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Adjust pH to 7.3.
Autoclave for 15 min at 15 psi pressure–121°C.
Vitamin B 12  Solution:
Composition per 10.0mL:
Vitamin B 12 .................................................................................1.0μg
Preparation of Vitamin B 12  Solution: Add vitamin B 12 to 10.0mL
of distilled/deionized water. Mix thoroughly. Filter sterilize.
Preparation of Medium: Aseptically mix 50.0mL of the sterile
agar solution with 50.0mL of the sterile mineral base, 2X. Aseptically
combine sterile solution T and sterile vitamin B 12 solution with the
sterile mineral base. Filter sterilize methanol and add to basal medium.
Pour into sterile Petri dishes or distribute into sterile tubes.
Use:  For the cultivation and maintenance of Methylophaga marina.