Part:BBa_K1895006:Experience

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The cells containing the BBa_K1895006 device were grown up in liquid culture of LB broth (with chloramphenicol in strains containing a pSB1C3 housed construct) overnight at 37°C. The following day, the bacterial cells were diluted down to an appropriate optical density in the region of 0.05 at 600 nm using LB broth with 0.034mg/ml chloramphenicol.

The plate reader was then set at either 30°C, 37°C and 42 °C and measured for growth using OD600 and fluorescence of GFP using 485 nm excitation wavelength and 520 nm emission wavelength with a gain of 1000.

The cells were left to grow for 18 hours with OD and fluorescence being measured every twenty minutes, and orbital shaking occuring between these measurement points. Our results can be seen below in Figure 1.

Figure 1. Summary of the data of all plate reader experiments. It can be seen that temperature does indeed affect the activity of both the P_htpGBBa_K1895000 and P_dnaKBBa_K1895006 promoters, thus leading to increasing levels of sfGFP expression. We have annotated the three states ('OFF', 'LOW' and 'HIGH') at which our 'light bulb' devices exist according to our three temperature conditions tested.

To see our results in more detail, check out the 2016 Newcastle University iGEM teams wiki