Selection of control genes for quantitative RT-PCR based on microarray data

Selection of control genes for quantitative RT-PCR based on microarray data

Author Shulzhenko, N. Google Scholar
Yambartsev, A. Google Scholar
Goncalves-Primo, A. Google Scholar
Gerbase-DeLima, M. Google Scholar
Morgun, A. Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Universidade de São Paulo (USP)
Abstract Use of internal reference gene(s) is necessary for adequate quantification of target gene expression by RT-PCR. Herein, we elaborated a strategy of control gene selection based on microarray data and illustrated it by analyzing endomyocardial biopsies with acute cardiac rejection and infection. Using order statistics and binomial distribution we evaluated the probability of finding low-varying genes by chance. for analysis, the microarray data were divided into two sample subsets. Among the first 10% of genes with the lowest standard deviations, we found 14 genes common to both subsets. After normalization using two selected genes, high correlation was observed between expression of target genes evaluated by microarray and RT-PCR, and in independent dataset by RT-PCR (r = 0.9, p < 0.001). in conclusion, we showed a simple and reliable strategy of selection and validation of control genes for RT-PCR from microarray data that can be easily applied for different experimental designs and tissues. (c) 2005 Elsevier Inc. All rights reserved.
Keywords normalization
RT-PCR
microarray
housekeeping gene
internal reference genes
heart transplant
Language English
Date 2005-11-11
Published in Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 337, n. 1, p. 306-312, 2005.
ISSN 0006-291X (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 306-312
Origin http://dx.doi.org/10.1016/j.bbrc.2005.09.048
Access rights Closed access
Type Article
Web of Science ID WOS:000232584000045
URI http://repositorio.unifesp.br/handle/11600/28549

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