Team:Osaka/Protocols

Protocols

Cell survival assay 1: UV irradiation

1. Pre-culture transformed cells in 3ml of LB medium at 37°C for 16h.
2. Induce parts with IPTG addition (to final concentration of 100µM) for 1h.
3. Dilute pre-culture samples according to intended UV energy dosage (if dosage is high, lower dilution rate is needed as fewer cells are expected to survive).
4. Transfer 50μl of diluted pre-culture onto LB agar plates and spread evenly. Air-dry the plates to remove excess wetness.
5. Irradiate cells on the agar with UV light at desired energy dosage.
6. Wrap plates in aluminium foil and incubate at 37°C.
7. After 16h, count number of colonies formed on control (non-irradiated) and UV-irradiated plates.