Team:Gifu/InterLab
InterLab
We joined InterLab Measurement Study in this year. We carried out all experiments according to iGEM 2018 InterLab Study Protocol and used "Safire ART-NR:F129013" as our plate reader.
Calibration 1: OD600Reference point - LUDOX
The Abs600 data of LUDOX and H2O is Table 1. And correction factor to transform our absorbance data into a standard OD600 measurement is calculated by the following formula.
Calibration 2: Particle Standard Curve - Microsphere
We prepared a dilution series of monodisperse silica microspheres and measure the Abs 600 in our plate reader. Using this measurement we construct a standard curve of particle concentration which can be used to convert Abs600 measurements to an estimated number of cells.
Calibration 3: Fluorescence standard curve - Fluorescein
We made a dilution series of fluorescein and measure the fluorescence in a 96 well plate in our plate reader. By measuring, we generate a standard curve of fluorescence for fluorescein concentration to convert your cell based readings to an equivalent fluorescein concentration.
Cell measurement
We transformed Escherichia coli DH5α with these following plasmids (all in pSB1C3) and incubated them in liquid culutures. We diluted the cultures further to a target Abs600 of 0.02 and incubated them for 6 hours at 37℃. At each time point 0 hours and 6 hours we take there samples. Finely we measured Abs600 and fluorescence of our samples.
Table 2 and 3 are the measurements of fluorescence and Abs600. Converting the data to the referense OD600 and the estimated number of cells, we calculated the fluorescence per OD600 and the fluorescence per particle. Fig 5 is the graph of the fluorescence per OD. Fig 6 is the graph of the fluorescence per particle.
