b-Actin -- an unsuitable internal control for RT-PCR

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Title b-Actin -- an unsuitable internal control for RT-PCR
Author Selvey, Saxon; Thompson, E.W.; Matthaei, K.; Lea, Rodney Arthur; Irving, Michael George; Griffiths, Lyn
Journal Name Molecular and Cellular Probes
Editor not stated
Year Published 2001
Place of publication UK
Publisher Academic Press
Abstract Despite reports confirming cell-cycle dependent gene expression and a number of studies describing specific circumstances in which β-actin is also regulated, the mRNA for β-actin remains a widely used housekeeping gene internal control. Utilizing differential reverse transcriptase-polymerase chain reaction (RT-PCR), we report here the dose-dependent inhibition of β-actin by matrigel. This was detected by comparison to the very moderate inhibition of the target gene, membrane type-1 matrix metalloproteinase (MT1-MMP), with results independently confirmed by similar findings on MT1-MMP expression using competitive RT-PCR. Furthermore, RT-PCR of the housekeeping gene 18 Svedberg Units (S) rRNA demonstrated excellent consistency, reproducibility and non-regulation by a matrigel treatment. We conclude that β-actin is highly regulated by matrigel and therefore unsuitable as an internal control in this treatment. Hence, these findings suggest that researchers have a responsibility to ensure that the housekeeping gene of choice is not regulated in their specific application, as such regulation may dramatically affect the accuracy of their results. This study reinforces the necessity for minimally regulated housekeeping genes such as 18S rRNA, and the superiority of competitive templates as internal controls for quantitative applications of RT-PCR.
Peer Reviewed Yes
Published Yes
Alternative URI http://dx.doi.org/10.1006/mcpr.2001.0376
Volume 15
Page from 307
Page to 311
ISSN 0890-8508
Date Accessioned 2002-06-24
Date Available 2015-05-06T21:37:03Z
Language en_US
Research Centre Menzies Health Institute Qld
Subject PRE2009-Medical and Health Sciences
URI http://hdl.handle.net/10072/3494
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1

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