Team:NYMU-Taipei/Experiments
Nitrogen Starvation
Culture Chlorella vulgaris
How long would it grow ?
- 7 days (theoretically)
- reach stationary phase at the fourth day (theoretically)
How to cultivate it ?
- Activate the algae in 15cc BG11(no need to unfreeze, keep it at the original took-out temperature) at 28°C~30°C
- Illumination at 10000~12000 lumen
How to regenerate it ?
- Each regenerate ’s plate put in 1500 microliter fully developed algae and 15cc BG11 (refill BG11 into the original plate)
- make sure the plate maintain in the needed amount
- regenerate every 7days
What we should do in these 7 days ?
- Experiment about nitrogen starvation
- Measurement about the yield of oil in the algae
Time guideline of nitrogen starvation experiment
Day 1
- Cultivate the algae and start doing the OD measurement every 6 hours (detailed protocol)
Day 2
- PCR , cloning and ligation (detailed protocol)
- Transformation and selection (detailed protocol)
Day 3
- Colony PCR and RE check (detailed protocol)
- DNA storage (detailed protocol)
Day 4
- Take some E.coli and ensure it could process nitrogen starvation by using kit
Day 5
- choose the successfully transformed e.coli and put it into the algae plate(when it reach its max od value) (detailed protocol)
- Start doing the measurement about oil production every 12 hours until you have at least 6 data (detailed protocol)
Day 6
- Keep on going the measurement
Day 7
- Keep on going the measurement
- Regenerate the algae
Day 8
- Back to day 1
*** this experiment group should have at least 4 routines *** those who incharge in the experiment should have wrote down the time and date *** once you have an extremely success-transformed E.coli, congratulate, you have your own free time in day 2 and day 3, but make sure you have done you liquid culture cultivation of e.coli so that you can continue your experiment at day 4
