Publication: MARIS: Method for Analyzing RNA following Intracellular Sorting
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Date
2014
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Public Library of Science
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Citation
Hrvatin, Siniša, Francis Deng, Charles W. O'Donnell, David K. Gifford, and Douglas A. Melton. 2014. “MARIS: Method for Analyzing RNA following Intracellular Sorting.” PLoS ONE 9 (3): e89459. doi:10.1371/journal.pone.0089459. http://dx.doi.org/10.1371/journal.pone.0089459.
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Abstract
Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human β cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines.
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Keywords
Biology, Anatomy and Physiology, Endocrine System, Endocrine Physiology, Endocrine Cells, Insulin, Computational Biology, Microarrays, Developmental Biology, Stem Cells, Embryonic Stem Cells, Cell Differentiation, Genomics, Genome Expression Analysis, Molecular cell biology, Gene expression, DNA transcription, Nucleic acids, RNA, RNA stability
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