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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

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Related Experiment Video

Updated: May 25, 2026

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Published on: October 27, 2011

Comparison between Normalised and Unnormalised 454-Sequencing Libraries for Small-Scale RNA-Seq Studies.

Robert Ekblom¹, Jon Slate, Gavin J Horsburgh

¹Department of Ecology and Genetics, Uppsala University, Norbyvägen 18 D, 75236 Uppsala, Sweden.

Comparative and Functional Genomics

|February 10, 2012

Summary

This summary is machine-generated.

Normalizing cDNA libraries before RNA-Seq improves gene detection and microsatellite discovery in nonmodel organisms like the zebra finch. This method enhances transcriptomic studies and is suitable for quantitative gene expression analysis.

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Last Updated: May 25, 2026

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

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Published on: October 27, 2011

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Area of Science:

Comparative genomics
Molecular biology
Bioinformatics

Background:

Next-generation sequencing of transcriptomes (RNA-Seq) is crucial for studying gene expression in nonmodel organisms.
Optimizing RNA-Seq library preparation is essential for maximizing data yield and accuracy.

Purpose of the Study:

To evaluate the effectiveness of normalizing complementary DNA (cDNA) libraries prior to sequencing in RNA-Seq studies.
To compare assembly quality, gene detection, and microsatellite discovery between normalized and unnormalized cDNA libraries in the zebra finch.

Main Methods:

RNA was extracted from zebra finch blood and spleen tissues.
Complementary DNA (cDNA) libraries were prepared, with one set normalized and the other unnormalized.
Libraries were sequenced using next-generation sequencing.
Transcriptome assemblies were generated and analyzed for contig number, read usage, gene detection, and microsatellite discovery.

Main Results:

Normalized libraries yielded assemblies with more contigs but used fewer reads compared to unnormalized libraries.
Significantly more genes were detected using contigs from normalized cDNA libraries.
Microsatellite discovery efficiency increased by up to 73% with normalized libraries.
Gene expression levels showed a positive correlation between normalized and unnormalized libraries.
No significant difference was observed in the number of differentially expressed genes between tissues for both library types.

Conclusions:

Normalized cDNA libraries are advantageous for various RNA-Seq applications, particularly in nonmodel organisms.
Normalization enhances gene discovery and marker identification efficiency.
Normalized libraries are suitable for reliable quantitative gene expression analysis, including differential expression studies.