PUBLICATION
Sample Preparation and Analysis of RNASeq-based Gene Expression Data from Zebrafish
- Authors
- Hostelley, T.L., Nesmith, J.E., Zaghloul, N.A.
- ID
- ZDB-PUB-171121-10
- Date
- 2017
- Source
- Journal of visualized experiments : JoVE (128): (Journal)
- Registered Authors
- Zaghloul, Norann A.
- Keywords
- none
- MeSH Terms
-
- Animals
- Gene Expression Profiling/methods*
- RNA/chemistry
- RNA/genetics*
- Sequence Analysis, RNA/methods*
- Zebrafish
- PubMed
- 29155729 Full text @ J. Vis. Exp.
Citation
Hostelley, T.L., Nesmith, J.E., Zaghloul, N.A. (2017) Sample Preparation and Analysis of RNASeq-based Gene Expression Data from Zebrafish. Journal of visualized experiments : JoVE. (128).
Abstract
The analysis of global gene expression changes is a valuable tool for identifying novel pathways underlying observed phenotypes. The zebrafish is an excellent model for rapid assessment of whole transcriptome from whole animal or individual cell populations due to the ease of isolation of RNA from large numbers of animals. Here a protocol for global gene expression analysis in zebrafish embryos using RNA sequencing (RNASeq) is presented. We describe preparation of RNA from whole embryos or from cell populations obtained using cell sorting in transgenic animals. We also describe an approach for analysis of RNASeq data to identify enriched pathways and Gene Ontology (GO) terms in global gene expression data sets. Finally, we provide a protocol for validation of gene expression changes using quantitative reverse transcriptase PCR (qRT-PCR). These protocols can be used for comparative analysis of control and experimental sets of zebrafish to identify novel gene expression changes, and provide molecular insight into phenotypes of interest.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping