Gen5 version required: 1.0 or higher
Basis for the Assay:
Fluorescent proteins are used as traceable genetic elements for a number of different cellular experiments. Fluorescent proteins are peptides that are inherently fluorescent and do not require any cofactors or modifications. EGFP is the brightest and most commonly used of the fluorescent proteins. EGFP is a variant that has had its genetic sequence modified to have excitation and emission wavelengths closer to that of fluorescein, as well as reflect human cells’ codon usage bias. EGFP, when translated, has an excitation and emission maxima of 488 nm and 509 nm respectively. Gene expression, in particular, is most easily performed in the microplate format.
The protocol calls for a fluorescence measurement using a 485/20 excitation filter and a 528/20 emission filter. Because EGFP experiments usually employ cells, fluorescent measurements are made from the bottom.
The experiment data file does not contain any data.
Plate configuration assumes that samples will be run in duplicate. The plate includes two blank wells, but no standards or controls wells. Thus, 47 samples can be run on a microplate in duplicate. The only data reduction performed is blank subtraction.
The report is configured to provide (1) blanked fluorescence.