RNA sequencing (RNA-Seq) technology enables rapid profiling and deep investigation of the transcriptome, for any species. This approach offers a number of advantages compared to microarray analysis, a legacy technology often used in gene expression studies.
Ability to detect novel transcripts: Unlike arrays, RNA-Seq technology does not require species- or transcript-specific probes. It can detect novel transcripts, gene fusions, single nucleotide variants, indels (small insertions and deletions), and other previously unknown changes that arrays cannot detect.1,2
Wider dynamic range: With array hybridization technology, gene expression measurement is limited by background at the low end and signal saturation at the high end. RNA-Seq technology produces discrete, digital sequencing read counts, and can quantify expression across a larger dynamic range (>105 for RNA-Seq vs. 103 for arrays).1,2,3
Higher specificity and sensitivity: Compared to microarrays, RNA-Seq technology can detect a higher percentage of differentially expressed genes, especially genes with low expression.4-6
Simple detection of rare and low-abundance transcripts: Sequencing coverage depth can easily be increased to detect rare transcripts, single transcripts per cell, or weakly expressed genes.
In the past, next-generation sequencing (NGS) data analysis required extensive bioinformatics expertise, presenting a major hurdle to adoption of RNA sequencing technology by biologists. The latest user-friendly tools vastly simplify the analysis process, providing accessible solutions for researchers without a bioinformatics background.
The portion of NIH grant funding allocated to new RNA sequencing vs. gene expression microarray-inclusive grants has been trending towards RNA-Seq technology for the last several years, and now constitutes the majority. Download our transcriptomics eBook to see the evidence.