Medium data

Search for medium no. = [744].

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

744 MODIFIED HB-1 MEDIUM

NaCl	`20.0`	g
MgSO₄·7H₂O	`3.5`	g
MgCl₂·6H₂O	`2.75`	g
KCl	`0.325`	g
NaBr	`0.05`	g
H₃BO₃	`0.015`	g
SrCl₂·6H₂O	`7.5`	mg
(NH₄)₂SO₄	`10.0`	mg
KI	`0.5`	mg
Na₂WO₄·2H₂O	`0.1`	mg
CaCl₂·2H₂O	`0.75`	g
KH₂PO₄	`0.5`	g
NiCl₂·6H₂O	`2.0`	mg
Trace minerals (see Medium No. 151	`10.0`	ml
Resazurin	`1.0`	mg
Distilled water	`1.0`	L

Mix ingredients, bring to a boil and cool under a H₂-CO₂ (4:1, v/v) gas mixture. Slowly add 10 ml 5% Na₂S·9H₂O solution to the medium, wait for the medium to become colorless, then adjust pH to 7.0 with NaOH. Distribute medium into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. Before inoculation, aseptically and anaerobically add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N₂ atmosphere):

10% KNO₃ solution	`10.0`	ml
0.05 M PIPES (pH 7.0) solution	`25.0`	ml
3% Na₂S·9H₂O solution (neutralized)	`4.0`	ml

Pressurize the inoculated culture vessels to 200 kPa H₂-CO₂ (4:1, v/v).