Endy:Victor3 Calculating fluorescent protein synthesis
Subtract a media background,, from the raw absorbance data, , and assume that the resulting data, , is directly proportional to the number of cells in the well.
Subtract a fluorescent protein-free cell background,, from the the raw fluorescent data, , and assume that the resulting data is proportional to the total number of GFP molecules in the well [immature GFP?].
Use standard calibration curves (see here for absorbance and here for fluorescence) to convert the background-corrected data into absolute units (CFU/well and GFP molecules per well). The calibration equations used are shown in Equations 3 & 4.
GFP synthesis rate calculations
To calculate the mean synthesis rate of GFP per cell,, assume the total GFP synthesis rate is equal to the time differential of . can be calculated as the total synthesis rate divided by .