An RNA-Seq Protocol for Differential Expression Analysis.
Cold Spring Harb Protoc
; 2019(6)2019 06 03.
Article
em En
| MEDLINE
| ID: mdl-30952685
ABSTRACT
Here we consider RNA-Seq, used to measure global gene expression through RNA fragmentation, capture, sequencing, and subsequent computational analysis. Xenopus, with its large number of RNA-rich, synchronously developing, and accessible embryos, is an excellent model organism for exploiting the power of high-throughput sequencing to understand gene expression during development. Here we present a standard RNA-Seq protocol for performing two-state differential gene expression analysis (between groups of replicates of control and treated embryos) using Illumina sequencing. Samples contain multiple whole embryos, and polyadenylated mRNA is measured under relative normalization. The protocol is divided into two parts wet-lab processes to prepare samples for sequencing and downstream computational analysis including quality control, quantification of gene expression, and differential expression.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Perfilação da Expressão Gênica
/
RNA-Seq
Limite:
Animals
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article