Please use this identifier to cite or link to this item: https://doi.org/10.2144/000114019
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dc.titleExpression analysis of rare cellular subsets: Direct RT-PCR on limited cell numbers obtained by FACS or soft agar assays
dc.contributor.authorHo, V.
dc.contributor.authorYeo, S.Y.
dc.contributor.authorKunasegaran, K.
dc.contributor.authorDe Silva, D.
dc.contributor.authorTarulli, G.A.
dc.contributor.authorVoorhoeve, P.M.
dc.contributor.authorPietersen, A.M.
dc.date.accessioned2014-11-26T08:28:16Z
dc.date.available2014-11-26T08:28:16Z
dc.date.issued2013-04
dc.identifier.citationHo, V., Yeo, S.Y., Kunasegaran, K., De Silva, D., Tarulli, G.A., Voorhoeve, P.M., Pietersen, A.M. (2013-04). Expression analysis of rare cellular subsets: Direct RT-PCR on limited cell numbers obtained by FACS or soft agar assays. BioTechniques 54 (4) : 208-211. ScholarBank@NUS Repository. https://doi.org/10.2144/000114019
dc.identifier.issn07366205
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/110076
dc.description.abstractSince tissues and tumors are heterogenous populations containing different cell types, their transcriptomes are blends of multiple mRNA expression profiles. Although fluorescence-activated cell sorting (FACS) allows isolation of individual cell types, RNA isolation and quantification remain problematic from rare subsets, such as tissue stem cells. Likewise, identification of transcriptional changes relevant to the tumorigenic potential of mammalian cells while they are actively growing as colonies in soft agar is also hampered by limited amounts of starting material. Here we describe a convenient method that fills the gap between single cell and whole tissue mRNA analysis, enabling mRNA quantification for individual colonies picked from soft agar. Our method involves directlysis, reverse transcription and quantitative PCR (RT-qPCR) on 500 sorted cells or a single soft agar colony, thus allowing evaluation of up to 20 transcripts in functionally distinct subpopulations without the need for RNA isolation or amplification.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.2144/000114019
dc.sourceScopus
dc.subjectFACS
dc.subjectGene expression profiling
dc.subjectRT-QPCR
dc.subjectSoft agar assay
dc.typeArticle
dc.contributor.departmentDUKE-NUS GRADUATE MEDICAL SCHOOL S'PORE
dc.description.doi10.2144/000114019
dc.description.sourcetitleBioTechniques
dc.description.volume54
dc.description.issue4
dc.description.page208-211
dc.description.codenBTNQD
dc.identifier.isiut000317712200006
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