title: RNA expression profiling from FACS-isolated cells of the Drosophila intestine.
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.
authors: DKFZ-ZMBH Alliance, University of Heidelberg, Heidelberg, Germany.
authors: Devanjali
authors: D
authors: Dutta
authors: DKFZ-ZMBH Alliance, University of Heidelberg, Heidelberg, Germany.
authors: Jinyi
authors: J
authors: Xiang
authors: DKFZ-ZMBH Alliance, University of Heidelberg, Heidelberg, Germany.
authors: Bruce A
authors: BA
authors: Edgar
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000166:cytology* / Q000235:genetics*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D007422:Intestines / Q000166:cytology*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D012313:RNA
keywords: Drosophila intestinal stem cells; cell type-specific RNA isolation; enterocyte; fluorescent-activated cell sorting (FACS); transcriptome profiling/RNASeq