UNIT 2F.2 RNA Expression Profiling from FACS‐Isolated Cells of the Drosophila Intestine. (13th November 2013)

• Record Type: Journal Article
• Authors: Dutta, Devanjali; Xiang, Jinyi; Edgar, Bruce A.
• Is Part Of: Current protocols in stem cell biology. Volume 27(2013)
• Page Start: 2F.2.1
• Page End: 2F.2.12
• Publication Date: 2013-11-13
• Subjects: Drosophila intestinal stem cells -- fluorescent‐activated cell sorting (FACS) -- cell type–specific RNA isolation -- transcriptome profiling/RNASeq -- enterocyte; Stem cells -- Laboratory manuals; Stem cells -- Physiology; Stem Cells; Stem cells; Laboratory Manual; Laboratory manuals; 571.5
• DOI: 10.1002/9780470151808.sc02f02s27
• Languages: English
• ISSNs: 1941-7322

Abstract

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.