Medium data

Search for medium no. = [850].

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

850 THERMOSYNTROPHA L-60 MEDIUM

Solution A:

KH₂PO₄	`0.54`	g
Na₂HPO₄	`1.33`	g
Yeast extract (BD-Difco)	`3.0`	g
Tryptone (BD-Difco)	`0.5`	g
Bacto peptone (BD-Difco)	`1.5`	g
Resazurin	`1.0`	mg
Distilled water	`760.0`	ml

Solution B:

NH₄Cl	`0.3`	g
NaCl	`0.3`	g
CaCl₂·2H₂O	`0.01`	g
MgCl₂·6H₂O	`0.1`	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:

Crotonic acid	`1.72`	g
Distilled water	`100.0`	ml

Adjust pH to 7.0.
Autoclave Solutions A, B and C separately. After cooling, aseptically and anaerobically 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 (use 5% solutions autoclaved and stored under a N₂ atmosphere).