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Specificity and dynamic range of detection with amplification

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posted on 30.12.2011 by Jorge M. Naciff, Brian D. Richardson, Kerry G. Oliver, M. Lynn Jump, Suzanne M. Torontali, Kenton D. Juhlin, Gregory J. Carr, Jennifer R. Paine, Jay P. Tiesman, George P. Daston

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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 2005


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.

Signal amplification does not change the linearity of the relationship between the amount of target quantified and the magnitude of the signal, even when the analyte is found within a complex mixture of cRNA potential targets. () Antisense oligonucleotide for , , , and at various concentrations (1, 10, 100, 1000, 10,000, and 100,000 amol/sample) were mixed and hybridized to the specific -, -, -, and -specific microspheres in a four-plex format, and the mean fluorescence intensity of at least 100 microspheres was determined after the hybridization signal was amplified as indicated in methods. () The indicated amounts of antisense oligonucleotide were mixed with fragmentation buffer or with various amounts of a highly complex cRNA sample, and then hybridized with -specific microspheres.