Assessment of normalization strategies for quantitative RT-PCR using microdissected tissue samples

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dc.contributor.authorErickson, Heidi S.
dc.contributor.authorAlbert, Paul S.
dc.contributor.authorGillespie, John W.
dc.contributor.authorWallis, Benjamin S.
dc.contributor.authorRodriguez-Canales, Jaime
dc.contributor.authorLinehan, W. Marston
dc.contributor.authorGonzalez, Sergio
dc.contributor.authorVelasco, Alfredo
dc.contributor.authorChuaqui, Rodrigo F.
dc.contributor.authorEmmert-Buck, Michael R.
dc.date.accessioned2025-01-21T01:05:26Z
dc.date.available2025-01-21T01:05:26Z
dc.date.issued2007
dc.description.abstractGene expression measurement techniques such as quantitative reverse transcriptase (qRT)-PCR require a normalization strategy to allow meaningful comparisons across biological samples. Typically, this is accomplished through the use of an endogenous housekeeping gene that is presumed to show stable expression levels in the samples under study. There is concern regarding how precisely specific genes can be measured in limited amounts of mRNA such as those from microdissected (MD) tissues. To address this issue, we evaluated three different approaches for qRT-PCR normalization of dissected samples; cell count during microdissection, total RNA measurement, and endogenous control genes. The data indicate that both cell count and total RNA are useful in calibrating input amounts at the outset of a study, but do not provide enough precision to serve as normalization standards. However, endogenous control genes can accurately determine the relative abundance of a target gene relative to the entire cellular transcriptome. Taken together, these results suggest that precise gene expression measurements can be made from MD samples if the appropriate normalization strategy is employed.
dc.description.funderIntramural NIH HHS
dc.fuente.origenWOS
dc.identifier.doi10.1038/labinvest.3700659
dc.identifier.eissn1530-0307
dc.identifier.issn0023-6837
dc.identifier.urihttps://doi.org/10.1038/labinvest.3700659
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/95920
dc.identifier.wosidWOS:000248943900010
dc.issue.numero9
dc.language.isoen
dc.pagina.final962
dc.pagina.inicio951
dc.revistaLaboratory investigation
dc.rightsacceso restringido
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleAssessment of normalization strategies for quantitative RT-PCR using microdissected tissue samples
dc.typeartículo
dc.volumen87
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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