Medium data

Search for medium no. = [636].

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

636 METHANOLOBUS R15 MEDIUM

Solution A:

KH₂PO₄	`0.54`	g
Na₂HPO₄	`1.34`	g
Yeast extract (BD-Difco)	`3.0`	g
Tryptone (BD-Difco)	`0.5`	g
Resazurin	`1.0`	mg
Distilled water	`860.0`	ml

Solution B:

NH₄Cl	`0.19`	g
NaCl	`0.19`	g
CaCl₂·2H₂O	`0.006`	g
MgCl₂·6H₂O	`0.063`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Distilled water	`100.0`	ml

Solution C:

50% Methanol 3.5 ml

Autoclave Solutions A and B separately and autoclave Solution C in a closed tube or vial. After cooling, combine the three solutions under a stream of N₂-CO₂ (80:20, v/v), then add the following solutions (filter-sterilized):

Trace vitamins (see Medium No. 197)	`10.0`	ml
8% NaHCO₃ solution	`30.0`	ml

Distribute the medium into culture vessels under the same gas mixture and seal with butyl rubber stoppers. Prior to inoculation, reduce the medium with 0.025% L-cysteine·HCl·H₂O and 0.025% Na₂S·9H₂O (autoclaved as 5% solutions under a N₂ atmosphere).