Influence of RT-qPCR primer position on EGFR interference efficacy in lung cancer cells

Biological Procedures Online

Research

Jacques De Grève¹  Erik Teugels¹  Gang Chen²  Peter Kronenberger³ 

[1] Laboratory of Molecular Oncology and Department of Medical Oncology, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Brussels, Belgium;Laboratory of Molecular Oncology and Department of Medical Oncology, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Brussels, Belgium;Department of Pathology, First Affiliated Hospital, Guangxi Medical University, Nanning Guangxi, PR, China;Laboratory of Molecular Oncology and Department of Medical Oncology, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Brussels, Belgium;Laboratory for Biotechnology, Department of Gezondheidszorg, Erasmushogeschool Brussel, Brussels, Belgium;

关键词: Epidermal Growth Factor Receptor;    H358 Cell;    Connective Tissue Growth Factor;    Epidermal Growth Factor Receptor Expression;    Epidermal Growth Factor Receptor Protein;   

DOI  :  10.1186/1480-9222-13-1

 received in 2010-05-29, accepted in 2010-10-20,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundReal-time quantitative RT-PCR (RT-qPCR) is a "gold" standard for measuring steady state mRNA levels in RNA interference assays. The knockdown of the epidermal growth factor receptor (EGFR) gene with eight individual EGFR small interfering RNAs (siRNAs) was estimated by RT-qPCR using three different RT-qPCR primer sets.ResultsOur results indicate that accurate measurement of siRNA efficacy by RT-qPCR requires careful attention for the selection of the primers used to amplify the target EGFR mRNA.ConclusionsWe conclude that when assessing siRNA efficacy with RT-qPCR, more than one primer set targeting different regions of the mRNA should be evaluated and at least one of these primer sets should amplify a region encompassing the siRNA recognition sequence.

【 授权许可】

Unknown   
© Chen et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.