Patent ID: 6351712
Filing Date: 2002-02-26
Classification: C12Q,G16B,Y10T

Abstract:
A method of fluorophore bias removal comprising the steps of:(a) labeling a first pool of genetic matter, derived from a biological system representing a baseline state, with a first fluorophore to obtain a first pool of fluorophore-labeled genetic matter; (b) labeling a second pool of genetic matter, derived from a biological system representing a perturbed state, with a second fluorophore to obtain a second pool of fluorophore-labeled genetic matter; (c) labeling a third pool of genetic matter, derived from said biological system representing said baseline state, with said second fluorophore to obtain a third pool of fluorophore-labeled genetic matter; (d) labeling a fourth pool of genetic matter, derived from said biological system representing said perturbed state, with said first fluorophore to obtain a fourth pool of fluorophore-labeled genetic matter; (e) contacting said first pool of fluorophore-labeled genetic matter and said second pool of fluorophore-labeled genetic matter with a first microarray under conditions such that hybridization can occur, respectively detecting a first and second flourescent emission signal at each of a plurality of discrete loci on the first microarray from said first and second pool of fluorophore-labeled genetic matter that is bound to said first microarray under said conditions, and determining a first color ratio between said first flourescent emission signal and said second flourescent emission signal; (f) contacting said third pool of fluorophore-labeled genetic matter and said fourth pool of fluorophore-labeled genetic matter with a second microarray under conditions such that hybridization can occur, respectively detecting a third and fourth flourescent emission signal at each of a plurality of discrete loci on the second microarray from said third and fourth pool of fluorophore-labeled genetic matter that is bound to said second microarray under said conditions, and determining a second color ratio between said third flourescent emission signal and said fourth flourescent emission signal; and (g) computing an average color ratio by averaging said first color ratio and said second color ratio; wherein: said first microarray and said second microarray are similar to each other, exact replicas of each other, or are identical; and said first color ratio and said second color ratio are determined for the same individual component of genetic matter, thereby removing fluorophore bias.