Comparison of the quantification of expression with and without signal amplification

Copyright information:Taken from "Design of a Microsphere-Based High-Throughput Gene Expression Assay to Determine Estrogenic Potential"Environmental Health Perspectives 2005;113(9):1164-1171.Published online 12 May 2005PMCID:PMC1280396.This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original DOI. Biotinylated cRNA samples from control (C) vehicle-treated or EE-exposed tissues (0.1, 1.0, or 10.0 μg/kg/day; L, M, and H, respectively) were hybridized to microspheres coupled to the specific probe for and the fluorescent intensity of at least 100 microspheres was determined (nonamplified). To increase the signal value, two equivalent sets of samples were hybridized to the specific microspheres, washed, and then hybridization signal was amplified as indicated in ”Materials and Methods.” A clear dose response to EE exposure can be determined in both cases; however, the amplification step increases the sensitivity of the assay.