Medium data

Search for medium no. = [594].

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

594 ALICYCLIPHILUS DENITRIFICANS MEDIUM

Solution A:

Na₂HPO₄·2H₂O	`0.53`	g
KH₂PO₄	`0.41`	g
NaNO3	`0.85`	g
Sodium acetate·3H₂O	`0.136`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Selenite-tungstate solution (see Medium No. 431)	`1.0`	ml
Distilled water	`900.0`	ml

Solution B:

CaCl₂2H₂O	`0.11`	g
MgCl₂·6H₂O	`0.1`	g
Distilled water	`50.0`	ml

Solution C:

NH₄HCO₃	`0.44`	g
NaHCO₃	`3.73`	g
Na₂SO₄	`0.2`	g
Distilled water	`50.0`	ml

Prepare all solutions and autoclave under a N₂ atmosphere. After cooling, combine the solutions and replace the gas phase with a N₂-CO₂ (4:1, v/v) gas mixture. Then, aseptically and anaerobically add the following autoclaved or *filter-sterilized solutions to the medium:

1.0 M Sodium acetate solution	`10.0`	ml
Vitamin solution* (see Medium No. 403)	`1.0`	ml
Thiamine solution* (see Medium No. 403)	`1.0`	ml
Vitamin B₁₂ solution* (see Medium No. 403)	`1.0`	ml

Check the final pH of the medium to be around 7.3.
Comment: JCM 14587 grows also in a modified medium with benzene and chlorate. For preparation of the modified medium, replace NaNO₃ with 1.06 g NaClO₃(in 900 ml solution A). After combining all solutions (A, B and C), add 1.0 ml/L of 1.0 M sodium acetate solution (autoclaved), 25 ml/L of 20 mM benzene (water-saturated, autoclaved), and 1 ml/L of each of vitamin solution (see Medium No. 403, filter-sterilized), thiamine solution (see Medium No. 403, filter-sterilized) and vitamin B₁₂ solution (see Medium No. 403, filter-sterilized) to the medium. Otherwise, prepare the modified medium as described above.