Name: R2A MEDIUM (MICROAEROBIC)
Creator: MediaDive
License: http://creativecommons.org/licenses/by/4.0/

Vol.

Main sol. 830d #

Equipment needed: Autoclave Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Yeast extract	0.50	g	0.5	-
Proteose peptone (BD DIFCO no. 3)	0.50	g	0.5	-
Casamino acids	0.50	g	0.5	-
Glucose	0.50	g	0.5	2.775
Starch , soluble	0.50	g	0.5	-
Na-pyruvate	0.30	g	0.3	2.726
K₂HPO₄	0.30	g	0.3	1.722
MgSO₄ x 7 H₂O	0.05	g	0.05	0.203
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients, then sparge solution with 80% N₂ and 20% CO₂ gas mixture for at least 30 - 45 min to remove dissolved oxygen and to saturate the solution with CO₂. Dispense solution in vials suitable for anaerobic cultures (e.g., Balch-type tubes or serum vials) to 30% volume (e.g., 5 ml medium in a 16 ml Hungate-type tube) under the 80% N₂ and 20% CO₂ gas atmosphere. Close vials with butyl rubber septa to prevent the free exchange of oxygen with the external atmosphere and autoclave them. Adjust pH of the autoclaved medium to 5.0 - 5.5 using a sterile stock solution of sodium carbonate (5% w/v), prepared under 80% N₂ and 20% CO₂ gas mixture.
2 Before inoculation, add sterile air to obtain 5% O₂ in headspace gas atmosphere (e.g., add 2 ml air to a Hungate-type tube).

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

All strains for this medium:	3
Strains with modifications:	1

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830d
830d . R2A MEDIUM (MICROAEROBIC)

830	R2A MEDIUM
a	MEDIUM R2A MODIFIED
b	MEDIUM R2A FOR DENITRIFYING BACTERIA
c	REACTIVATION WITH LIQUID MEDIUM 830
d	R2A MEDIUM (MICROAEROBIC)