| Genome-wide transcriptome analysis of 150 cell samples. | |
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MedLine Citation:
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PMID: 20023796 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A 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 in 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. |
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Authors:
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Daniel Irimia; Michael Mindrinos; Aman Russom; Wenzhong Xiao; Julie Wilhelmy; Shenglong Wang; Joe Don Heath; Nurith Kurn; Ronald G Tompkins; Ronald W Davis; Mehmet Toner |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural Date: 2008-11-12 |
Journal Detail:
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Title: Integrative biology : quantitative biosciences from nano to macro Volume: 1 ISSN: 1757-9708 ISO Abbreviation: Integr Biol (Camb) Publication Date: 2009 Jan |
Date Detail:
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Created Date: 2009-12-21 Completed Date: 2010-03-10 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101478378 Medline TA: Integr Biol (Camb) Country: England |
Other Details:
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Languages: eng Pagination: 99-107 Citation Subset: IM |
Affiliation:
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BioMEMS Resource Center, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, 02114, USA. dirimia@hms.harvard.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Cell Separation
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instrumentation*,
methods Chromosome Mapping / instrumentation*, methods Equipment Design Equipment Failure Analysis Gene Expression Profiling / instrumentation*, methods Microfluidic Analytical Techniques / instrumentation*, methods Nucleic Acid Amplification Techniques / instrumentation*, methods RNA / genetics*, isolation & purification |
| Grant Support | |
ID/Acronym/Agency:
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P01HG000205/HG/NHGRI NIH HHS; U54GM62119/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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63231-63-0/RNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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