Part:BBa_K1323010:Experience

The fluorescence and optical density of DH5α containing YFP was observed over time and compared to DH5α that did not contain YFP. Method: The expression cassette (in the pSB3K3 backbone) contained the YFP coding sequence under the constitutive sarA promoter (BBa_K1323021) and TIR ribosome binding site (BBa_K1323016). DH5α either containing the YFP plasmid or containing no plasmids were grown in triplicate overnight at 37°C in LB supplemented with the appropriate antibiotics. Cultures were diluted 1/100 into fresh LB with appropriate antibiotics and incubated at 37°C. Every hour, the fluorescence (excitation: 485 nm, emission: 535 nm) and optical density at 600 nm of 200 μl of culture was read using a plate reader. Conclusion: The YFP coding sequence is able to produce a fluorescent protein.

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