Angie L. Bookout and David J. Mangelsdorf
Corresponding Author:
davo.mango@utsouthwestern.eduHoward Hughes Medical Institute, Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX, USA A major goal of the Nuclear Receptor Signaling Atlas (NURSA) is to elucidate the biochemical and physiological roles of nuclear receptors in vivo. Characterizing the tissue expression pattern of individual receptors and their target genes in whole animals under various pharmacological conditions and genotypes is an essential component of this aim. Here we describe a high-throughput quantitative, real-time, reverse-transcription PCR (QPCR) method for the measurement of both the relative level of expression of a particular transcript in a given tissue or cell type, and the relative change in expression of a particular transcript after pharmacologic or genotypic manipulation.This method is provided as a standardized protocol for those in the nuclear receptor field. It is meant to be a simplified, easy to use protocol for the rapid, high-throughput measurement of transcript levels in a large number of samples. A subsequent report will provide validated primer and probe sequence information for the entire mouse and human nuclear receptor superfamily.
Received Novemeber 4th, 2003; Accepted December 12th, 2003; Published December 23rd, 2003 | Abbreviations: GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; GOI: Gene of interest; GSRP: Gene-specific reverse primer; PCR: polymerase chain reaction; QPCR: Quantitative real-time PCR; Tm: Amplicon melting temperature | Copyright © 2003, Bookout and Mangelsdorf. This is an open-access article distributed under the terms of the Creative Commons Non-Commercial Attribution License, which permits unrestricted non-commercial use distribution and reproduction in any medium, provided the original work is properly cited.
Cite this article: Nuclear Receptor Signaling (2003) 1, e012
