Medium data

Search for medium no. = [1365].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1365 ACIDIC LOW SALT MINERAL MEDIUM

MgSO₄·7H₂O	`98.6`	mg
CaCl₂·2H₂O	`14.7`	mg
K₂SO₄	`34.9`	mg
(NH₄)₂SO₄	`0.264`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Selenite-tungustate solution (see below)	`1.0`	ml
Distilled water	`1.0`	L

Mix components thoroughly. For preparation of solid medium, add 15.0 g/L agarose. Autoclave and, after cooling to 60C, add the following solutions (autoclaved or *filter-sterilized):

0.1 M KH₂PO₄ solution	`1.0`	ml
Trace vitamins* (see Medium No. 197)	`10.0`	ml
Vitamin B₁₂ solution* (see Medium No. 403)	`4.0`	ml
1.0 mM CeCl₃ solution*	`1.0`	ml
1.0 N H₂SO₄ solution*	`4.0`	ml

Check pH to be 2.0 to 3.0. Cultivate with methane or methanol as a substrate as described below:
Methane. Aseptically distribute the medium into sterile serum bottles (leave ≥ 80% gas phase by volume) and seal with rubber stoppers. Add sterile CO₂ and methane (respectively 2-10% and 5-20% by volume) in the gas phase. Cultivate with shaking.
Methanol: Aseptically distribute the medium into culture vessels (e.g., 10 ml in 60 ml serum bottles), supplement with 0.4-1.0 ml per liter methanol (filter-sterilized) and seal with rubber stoppers. Add sterile CO₂ (2-10% by volume) in the gas phase.
Selenite-tungstate solution:

NaOH	`0.4`	g
Na₂SeO₃·5H₂O	`0.06`	g
Na2WO₄·2H₂O	`0.08`	g
Distilled water	`1.0`	L