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Media Information

Media No. 402 Methylophaga medium

Composition

capacity

unit

Vitamin B₁₂ (0.1 mcg/ml) filter-sterilized

1.0

Methanol, filter-sterilized

0.3

For solid medium prepare a 2X Mineral Base solution and a 3% agar solution and autoclave both separately. Cool to 50℃ and aseptically mix the solutions in equal volume before adding solution T, the B_12 _solution and methanol.

Solution T:

Composition	capacity	unit
NH₄Cl	10.0	g
Bis-Tris	10.0	g
Ferric ammonium citrate	0.3	g
Distilled water	100.0	ml
Adjust to pH 7.3. Autoclave at 121\|·\| for 15 minutes.
KH₂PO₄	0.7	g

Mineral Base:

Composition	capacity	unit
NaCl	24.0	g
KCl	0.5	g
Bis-Tris	0.5	g
Wolfe's Mineral Solution (see below)	10.0	ml
Distilled water	990.0	ml
Adjust to pH 7.3 with HCl. Autoclave at 121\|·\| for 15 minutes.
MgCl₂ . 6H₂O	3.0	g
MgSO₄ . 7H₂O	2.0	g
CaCl₂ . 2H₂O	1.0	g

Complete Medium:

Wolfe's Mineral Solution:

Composition	capacity	unit
Nitrilotriacetic acid	1.5	g
NaCl	1.0	g
Distilled water	1.0	L
Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.
MgSO₄ . 7H₂O	3.0	g
MnSO₄ . H₂O	0.5	g
FeSO₄ . 7H₂O	0.1	g
CoCl₂ . 6H₂O	0.1	g
CaCl₂	0.1	g
ZnSO₄ . 7H₂O	0.1	g
CuSO₄ . 5H₂O	0.01	g
AlK(SO₄)₂ . 12H₂O	0.01	g
H₃BO₃	0.01	g
Na₂MoO₄ . 2H₂O	0.01	g

Aseptically combine the following solutions just before use:

Mineral Base (see below),

100.0

Solution T (see below)

2.0

** ph : -

** memo : -

** additional actions :

Korean Collection for Type Cultures (KCTC)
Korea Research Institute of Bioscience and Biotechnology (KRIBB)
181 Ipsin-gil. Jeongeup-si, Jeollabuk-do 56212, Korea

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