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dc.contributor.authorThompson, E.
dc.contributor.authorMatthaei, K.
dc.contributor.authorLea, Rodney
dc.contributor.authorIrving, Michael
dc.contributor.authorGriffiths, Lyn
dc.contributor.editornot stated
dc.date.accessioned2017-05-03T13:33:47Z
dc.date.available2017-05-03T13:33:47Z
dc.date.issued2001
dc.identifier.issn08908508
dc.identifier.doi10.1006/mcpr.2001.0376
dc.identifier.urihttp://hdl.handle.net/10072/3494
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.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherAcademic Press
dc.publisher.placeUK
dc.relation.ispartofpagefrom307
dc.relation.ispartofpageto311
dc.relation.ispartofjournalMolecular and Cellular Probes
dc.relation.ispartofvolume15
dc.subject.fieldofresearchMedicinal and biomolecular chemistry
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchcode3404
dc.subject.fieldofresearchcode3101
dc.titleb-Actin -- an unsuitable internal control for RT-PCR
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2015-05-06T21:37:03Z
gro.hasfulltextNo Full Text
gro.griffith.authorIrving, Michael G.
gro.griffith.authorGriffiths, Lyn
gro.griffith.authorSelvey, Saxon
gro.griffith.authorLea, Rodney A.


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