# Endy:Victor3 Calculating fluorescent protein synthesis

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Add to My Links## Background subtraction

Subtract a media background, *A*_{media}, from the raw absorbance data, *A*_{raw}, and assume that the resulting data, *A*_{corrected}, is directly proportional to the number of cells in the well.

...Equation 1 |

Subtract a fluorescent protein-free cell background, *G*_{cells}, from the the raw fluorescent data, *G*_{raw}, and assume that the resulting data *G*_{corrected} 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, *S*_{cell}, assume the total GFP synthesis rate is equal to the time differential of *G**F**P*. *S*_{cell} can be calculated as the total synthesis rate divided by *C**F**U*.

...Equation 5 | ||

...Equation 6 |