Part:BBa_K1968016:Experience

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Applications of BBa_K1968016

iLOV was characterized in E. coli TOP10 cells. Colonies were picked and grown overnight at 37 °C, 250 RPM in 50 mL conical tubes in 10 mL of LB medium with its respective antibiotic (50 ug/mL kanamycin). Cell density was measured in a Jenway 7305 spectrophotometer, the obtained data was processed using Excel Data Processing tools. Cells were then subcultured into opaque-walled 96 well plates and adjusted to OD600 0.05 in 290 µl of LB medium supplemented with antibiotics in . Plates were then placed in the Fluostar omega microplate reader from BMG labtech, and grown at 37 °C for 20 hours at 300 RPM. Aside from monitoring OD600, fluorescence was determined by measuring emission wavelengths using one excitation filter (480nm ) and emission filter (530nm) every 5 minutes. All data is presented in relative iLOV fluorescence units, which represents the relative amount of the specific wavelengths of light collected by the plate reader at the conditions described above.

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