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dc.contributor.authorThompson, E.en_US
dc.contributor.authorMatthaei, K.en_US
dc.contributor.authorLea, Rodneyen_US
dc.contributor.authorIrving, Michaelen_US
dc.contributor.authorGriffiths, Lynen_US
dc.contributor.editornot stateden_US
dc.description.abstractDespite reports confirming cell-cycle dependent gene expression and a number of studies describing specific circumstances in which ߭actin is also regulated, the mRNA for ߭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 ߭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 ߭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.en_US
dc.publisherAcademic Pressen_US
dc.relation.ispartofjournalMolecular and Cellular Probesen_US
dc.titleb-Actin -- an unsuitable internal control for RT-PCRen_US
dc.typeJournal articleen_US
dc.type.descriptionC1 - Peer Reviewed (HERDC)en_US
dc.type.codeC - Journal Articlesen_US
gro.hasfulltextNo Full Text

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