Methods:UV Sensitivity Assay
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UV irradiation causes DNA damage that must be repaired in order for replication of the chromosome to occur. This assay is used to determine whether a strain's recombinational repair mechanisms have been compromised. Failure to repair the breaks will lead to induction of the SOS Response and a decrease in cell viability.
- UV light source
- Control Strains:
- Recombination proficient control (like MG1655)
- Recombination-deficient control (like DH5&alpha')
- Grow cells to A600 = 0.4
- Plate serial dilutions onto LB plates
- Grow for 30 minutes
- Expose plates to UV light
- Incubate overnight (up to 72 hours)
- Count colonies
- Protocol taken from Lopez et al. (2005)
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