Medium data

Search for medium no. = [1304].

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

1304 FRESHWATER MEDIUM WITH RHAMNOSE

NaCl	`1.0`	g
MgCl₂·6H₂O	`0.4`	g
CaCl₂·2H₂O	`0.15`	g
NH₄Cl	`0.25`	g
KH₂PO₄	`0.2`	g
KCl	`0.5`	g
Resazurin	`0.5`	mg
Distilled water	`955.0`	ml

Mix components thoroughly, adjust pH to 7.2-7.4 and autoclave under a N₂-CO₂ (80:20, v/v) gas mixture. After cooling, while gassing with the same gas mixture, add the following solutions from sterile anaerobic stocks (autoclaved or *filter-sterilized):

8% NaHCO₃ solution*	`30.0`	ml
Modified trace element mixture (see below)	`1.0`	ml
Vitamin solution* (see Medium No. 556)	`1.0`	ml
Selenite-tungstate solution (see Medium No. 431)	`0.5`	ml
0.5 M L-Rhamnose solution*	`10.0`	ml
10 % Yeast extract solution	`5.0`	ml

Readjust pH to 7.2-7.4, if necessary. Aseptically and anaerobically distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under the same gas mixture and seal with butyl rubber stoppers. Prior to use, add per liter the following solution (autoclaved and stored under N₂):

5% Na₂S·9H₂O solution 5.0 ml

Modified trace element mixture:

EDTA·Na₂	`5.2`	g
FeCl₂·4H₂O	`1.0`	g
H₃BO₃	`30.0`	mg
MnCl₂·4H₂O	`100.0`	mg
CoCl₂-6H₂O	`190.0`	mg
NiCl₂-6H₂O	`24.0`	mg
CuCl₂-2H₂O	`2.0`	mg
ZnSO₄-7H₂O	`144.0`	mg
Na₂MoO₄-2H₂O	`36.0`	mg
Distilled water	`1.0`	L

Dissolve EDTA·Na₂ in distilled water first and adjust pH to 6.5 with NaOH. Then, proceed to add minerals.