We use two type of medium for growing R. sphaeroides; Sistrom's minimal medium and LB (below).

Generally, Sistrom's minimal medium is used for all the work involving any biochemical/biophyscial measurements. This is the lab "default" medium. LB is generally only used for the preparation of genomic DNA, or, for the growth of auxotrophs.

Sistrom's medium is used for both liquid and plate culture. To make plates, agar is added to Sistrom's medium to 1.5% (w/v) then autoclaved.

Whether in liquid or solid media, we grow R. sphaeroides at 28-30 degrees Celsius. It takes 3-4 days for the formation of ~1mm diameter colonies.

When R. sphaeroides is grown on plates the colonies should look like those shown.

Liquid Cultures
 
 

Tryptone:                    10g
Yeast extract:               5g
NaCl:                          10g
 

Make the volume up to 800 ml. Adjust the pH to 7.0 then adjust final volume to 1 L.

For LB agar, add 15g of agar to before adjusting volume to 1 L.