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Genome-wide transcriptome analysis of 150 cell samples
Harvard Univ, Sch Med, Boston ; Massachusetts Gen Hosp, Shriners Hosp Children.
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2009 (English)In: Integrative Biology, ISSN 1757-9694, E-ISSN 1757-9708, Vol. 1, no 1, 99-107 p.Article in journal (Refereed) Published
Abstract [en]

 major challenge in molecular biology is interrogating the human transcriptome on a genome wide scale when only a limited amount of biological sample is available for analysis. Current methodologies using microarray technologies for simultaneously monitoring mRNA transcription levels, require nanogram amounts of total RNA. To overcome the sample size limitation of Current technologies, we have developed a method to probe the global gene expression ill biological samples as small as 150 cells, or the equivalent of approximately 300 pg total RNA. The new method employs microfluidic devices for the purification of total RNA from mammalian cells and ultra-sensitive whole transcriptome amplification techniques. We verified that the RNA integrity is preserved through the isolation process, accomplished highly reproducible whole transcriptome analysis, and established high correlation between repeated isolations of 150 cells and the same cell culture sample. We validated the technology by demonstrating that the combined microfluidic and amplification protocol is capable of identifying biological pathways perturbed by Stimulation, which are consistent with the information recognized in bulk-isolated samples.

Place, publisher, year, edition, pages
2009. Vol. 1, no 1, 99-107 p.
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Natural Sciences
Identifiers
URN: urn:nbn:se:kth:diva-89923DOI: 10.1039/b814329cISI: 000266978200011OAI: oai:DiVA.org:kth-89923DiVA: diva2:503990
Note
QC 20120219Available from: 2012-02-17 Created: 2012-02-17 Last updated: 2017-12-07Bibliographically approved

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