RNA expression profiling from FACS-isolated cells of the Drosophila intestine

• Verfasst von: Dutta, Devanjali [VerfasserIn]
• Verfasst von: Xiang, Jinyi [VerfasserIn]
• Verfasst von: Edgar, Bruce [VerfasserIn]
• Titel: RNA expression profiling from FACS-isolated cells of the Drosophila intestine
• Verf.angabe: Devanjali Dutta, Jinyi Xiang, Bruce A. Edgar
• E-Jahr: 2013
• Jahr: 13 November 2013
• Umfang: 12 S.
• Fussnoten: Gesehen am 05.01.2021
• Titel Quelle: Enthalten in: Current protocols in stem cell biology
• Ort Quelle: New York, NY [u.a.] : Wiley, 2007
• Jahr Quelle: 2013
• Band/Heft Quelle: 27(2013), 1, Seite 2F.2.1-2F.2.12
• ISSN Quelle: 1938-8969
• DOI: doi:10.1002/9780470151808.sc02f02s27
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: cell type-specific RNA isolation
• Sach-SW: Drosophila intestinal stem cells
• Sach-SW: enterocyte
• Sach-SW: fluorescent-activated cell sorting (FACS)
• Sach-SW: transcriptome profiling/RNASeq
• K10plus-PPN: 1743958544

Abstract

This unit describes a protocol for the isolation of Drosophila intestinal cell populations for the purpose of cell type-specific transcriptome profiling. A method to select a cell type of interest labeled with green or yellow fluorescent protein (GFP, YFP) by making use of the GAL4-UAS bipartite system and fluorescent-activated cell sorting (FACS) is presented. Total RNA is isolated from the sorted cells and linear RNA amplification is used to obtain sufficient amounts of high-quality RNA for analysis by microarray, RT-PCR, or RNA sequencing. This method will be useful for quantitative transcriptome comparison across intestinal cell types under normal and various experimental conditions. Curr. Protoc. Stem Cell Biol. 27:2F.2.1-2F.2.12. © 2013 by John Wiley & Sons, Inc.