Endy:Victor3 Calculating fluorescent protein synthesis

From OpenWetWare
Jump to: navigation, search

Background subtraction

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.

...Equation 1

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?].

...Equation 2

Unit conversion

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.

...Equation 3
...Equation 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 .

...Equation 5
...Equation 6