Medium data

Search for medium no. = [859].

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

859 PYROLOBUS MEDIUM

Solution A:

Synthetic seawater (2 x, see Medium No. 244)	`250.0`	ml
Wolfe's mineral elixier (see Medium No. 470)	`1.0`	ml
MES	`1.95`	g
Resazurin	`0.5`	mg
Distilled water	`750.0`	ml

Solution B:

KNO₃	`1.0`	g
KH₂PO₄	`0.5`	g
Distilled water	`10.0`	ml

Solution C:

Na₂S·9H₂O	`0.15`	g
Distilled water	`10.0`	ml

Mix components of solution A, adjust pH to 5.7, then boil for 1 min and cool down to room temperature under a stream of H₂-CO₂ (4:1, v/v) gas mixture. Dispense the medium into culture vessels (e.g., 20 ml in 120 ml serum bottles) under the same gas mixture, seal with butyl rubber stoppers and autoclave. Separately autoclave Solutions B and C under a N₂ atmosphere. Prior to inoculation, add Solutions B and C (10.0 ml each per liter of Solution A) and readjust pH to 5.7, if necessary. Pressurize inoculated bottles to 200 kPa H₂-CO₂ (4:1, v/v).