Tips for growing O. tauri, strain RCC745
  • Only use plastic spoons to handle ASW components
  • To suppress bacterial growth, add antibiotic mix (see page 3 of ASW recipe)
  • In our experience, cells at OD600 = 0.1 are approaching stationary phase
Here are a few pictures of our incubator setup, including details of parts/parameters.

We grow our cells at 24°C. In our experience, 20 - 24°C is fine. In the wild, these cells can tolerate lower winter temperatures, but probably grow slower. The incubator is a Binder KB115.

Our growth conditions:
Agitation: ~60 RPM, using a Thermo 2309-1CEQ rotator.
Light color: Lee #183, "moonlight blue"
Illuminance: ~320 lux, measured with the sensor on the shaker.
Light-dark cycle: 12 hours : 12 hours

The lights are white "Angel Eye" LEDs that are mounted around the headlights of sports cars. These LEDs are available in hardware stores. We use the Lee #183 filter, but other blue filters should also work. We've also been told (but haven't tried) that it's even easier to use blue LEDs.

The lighting can be controlled with a simple analog timer (blocky adapter with blue spokes), also available from hardware stores.

When cells are growing well in log phase under the 12h:12h light-dark regime, OD600 should double every 24 hours. This doubling only lasts until the cells reach OD600 ~ 0.1, at which point the culture is starting to enter stationary phase. To track growth more accurately, use flow cytometry.

Another way to check cell growth is by DIC light microscopy. Healthy cells should look nice and round, as seen on our research page. There should be no bacteria, which swim around rapidly. In comparison O. tauri 'wanders' around.

On the left is a photo of a late log-phase culture (
OD600 > 0.4). If this culture was contaminated with bacteria, the color would be off-white instead.