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Gene Expression Analysis of Sorted Cells by RNA-seq in Drosophila Intestine   

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Abstract

RNA sequencing (RNA-seq) has become a popular method for profiling gene expression. Among many applications, one common purpose is to identify differentially expressed genes and pathways in different biological or pathological conditions. This protocol provides detailed procedure for RNA-seq analysis of ~250,000 sorted Drosophila intestinal cells (Chen et al., 2016), in which RNA amplification is not required.

Keywords: RNA-seq, FACS, Drosophila intestine, Progenitor cells, Transcriptome analysis

Background

Transcriptome analysis by RNA-seq has become a popular method for the identification of differentially expressed genes and pathways under different biological or pathological conditions. For samples that yield low mRNA levels, RNA or cDNA amplification was commonly performed before deep-sequencing (Dutta et al., 2015). However, this procedure could potentially omit important candidates that are expressed in low abundance. Here we provide a detailed procedure for RNA-seq analysis of sorted Drosophila gut cells in which RNA amplification is not required.

Copyright Chen et al. This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0).
How to cite:  Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
  1. Chen, J., Li, J., Huang, H. and Xi, R. (2016). Gene Expression Analysis of Sorted Cells by RNA-seq in Drosophila Intestine. Bio-protocol 6(24): e2079. DOI: 10.21769/BioProtoc.2079.
  2. Chen, J., Xu, N., Huang, H., Cai, T. and Xi, R. (2016). A feedback amplification loop between stem cells and their progeny promotes tissue regeneration and tumorigenesis. Elife 5. pii: e14330
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