Team:Aix-Marseille/SN
SuperNova activity
We wanted to test the activity of [http://parts.igem.org/Part:BBa_K1491018 Supernova] on the colonies, so we made a protocol where we illuminated Petri dish with a 16,5 Candela 595nm Diode (lumex ssl-lx5093syc/g) for 2 hours.
Protocol:
- Transform 100µL TG1 with Eprc - SN (psb1C3 plasmid, 5µL)
- Into 2mL LB add 2µL of 2 DO TG1 (to get an adequate number of colonies)
- Pipet up and down
- From the first dilution, take 8µL and dilute it into 1mL LB
- Spin at 5000rpm for 10min
- Remove 850µL supernatant
- Resuspend pellet (even if you can't see it)
- Spread remaining 150µL on petri dish
- Immediatly put on 590nm light / put control in dark
- After two hours, incubate overnight
- Count colonies
| Exposed | Not exposed | |
| Supernova | 1496 | 3672 |
| RFP | 3545 | 3638 |
With a RFP plasmid as a control, we have shown that there's no significant decrease of the number of colonies when the Petri dish is exposed to the light compared to when bacteria are not exposed to the light.
With this experiment, we can conclude that supernova allows us to divide the number of colonies by 2,45. Thus, its toxic action is proved.
