Q: We are having some trouble with our cultures and were wondering if you would be able to provide any additional advice or information. Some questions that we have or have run across as we began to grow the algae are as follows:
* Have you used instant ocean to prepare your medium?
* What colors we should be seeing and should we be seeing any sedimentation?
- i.e. Is there a color change that would indicate the algae is unhealthy?
* Do you have any references or pointers as to what timescale and methods are best for splitting and scaling-up the cultures (e.g. what density should we grow to before we split and what splitting ratio works best with I. galbana).
* Should we grow these in vented flasks or tubes or should closed glass tubes work?
* For larger scale production, do you recommend bubbling air or 1% CO2?
A: 1. We do not use instant ocean. Instant ocean will precipitate when autoclaved which may make key nutrients unavailable.
2. CCMP1323 should be a nice light to golden brown when in a healthy culture. any other color such as green would indicate an unhealthy culture.
3. I like a 1 to 5 split for keeping a good growth rate and building biomass once a culture is healthy my co-worker prefers a 1 to 10 split. 1:1 is good if it is unhealthy. For this culture we put 1 drop into 18mls and it is grown in 3 weeks.
4. We use Kim caps on test tubes https://labgenome.com/73660-25-dwk-life-sciences-kimble-kim-kap-test-tube-caps-case-of-500/?gclid=EAIaIQobChMI7v2znd7b5wIViaztCh10ZgazEAQYASABEgLyyPD_BwE or silicone stoppers in a flask. Open or vented will allow bacteria in.
5. For volumes over 20L we sometimes use CO2.
6. I would not recommend agitation or shaking.