Medium data

Search for medium no. = [499].

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

499 SULFATE-FREE SYNTROPHOMONUS MEDIUM

Solution A:

Mineral solution (see below)	`50.0`	ml
Trace element solution SL-10 (see Medium No. 433)	`1.0`	ml
Rumen fluid, clarified (see Medium No. 266)	`50.0`	ml
Trypticase peptone (BD-BBL)	`1.0`	g
Vitamin solution (see below)	`5.0`	ml
Crotonic acid	`1.7`	g
Resazurin	`1.0`	mg
Distilled water	`810.0`	ml

Solution B:

NaHCO₃	`3.5`	g
Distilled water	`70.0`	ml

Solution C:

L-Cysteine·HCl·H₂O	`0.3`	g
Distilled water	`10.0`	ml

Solution D:

Na₂S·9H₂O	`0.3`	g
Distilled water	`10.0`	ml

Mix ingredients of Solution A, adjust pH to 7.2, bring to a boil for a few minutes and cool down to room temperature under a stream of N₂-CO₂ (4:1, v/v) gas mixture. Distribute the solution into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. Filter-sterilize Solution B and equilibrate by gassing with the same gas mixture for at least 15 min. Autoclave Solutions C and D under a N₂ atmosphere. Add appropriate amounts of Solutions B, C and D to Solution A to complete the medium.
Mineral solution:

KH₂PO₄	`10.0`	g
MgCl₂·6H₂O	`6.6`	g
NaCl	`8.0`	g
NH₄Cl	`8.0`	g
CaCl₂·2H₂O	`1.0`	g
Distilled water	`1.0`	L

Vitamin solution:

Biotin	`0.25`	mg
Nicotinic acid	`2.5`	mg
Thiamine·HCl	`1.25`	mg
p-Aminobenzoic acid	`1.25`	mg
Pantothenic acid	`0.62`	mg
Pyridoxine·HCl	`6.2`	mg
Distilled water	`1.0`	L