Rapid Culture

From ChoanoWiki

Roberta Hannibal 11/10/04

1. Find log phase of growth (for Monosiga, b/w 12-26hrs)

2. Pick convenient time period in log phase to split culture (for Monosiga, 24hrs)

3. Start with healthy culture, 10⁶-10⁷ cells/mL (for Monosiga, 3-4 days old culture)

4. Split culture every 24hrs (or whatever time period decided upon)

5. Repeat three more times, at least (less times may work also)

• I think it would be best to do this no longer than a week; if you need to repeat the experiment, its probably better to start again with a healthy, grown up culture

• For the last split, it may be good to split at a later time than usual (ie: let culture grow up more, than split so time between last split and time fixed is shorter). I had really good results 11/5/04 doing this, but it may have been because I examined my slide very closely.

ex: Monosiga (11/5/04)

starting culture: started -- 10/27/04

first split: -- 10/31/04 -- 10am

second split: -- 11/1/04 -- 12 am

third split: -- 11/2/04 -- 11:20am

forth split: -- 11/3/04 -- 11:10am

fifth (and last) split: -- 11/4/04 -- 7pm

cells fixed: -- 11/5/04 -- around10am