RNA sequencing (RNA-Seq) is a powerful tool with a multitude of applications, including:
- Gene expression profiling
- Whole transcriptome research
- Alternative splicing isoform identification
- Rare and novel transcript discovery
- Research on gene fusions, isoforms, and structural variants
- Allele-specific expression studies
- Research on disease-associated SNVs
- Total RNA + hybridization capture enrichment experiments
RNA-Seq has facilitated transcriptomics, which is defined as the study of the entire complement of RNA in a sample. Sequencing RNA provides a snapshot of the genes are expressed within a particular tissue, cell type, or even a single cell. Environmental changes that affect the whole organism or even the microenvironment of a single cell induce genes to turn on and off in response. Researching changes in gene expression provides a wealth of information that facilitates a better understanding of genomics, cellular regulation, and overall function of an organism.
RNA-Seq strategies use NGS technology to understand the transcriptome and provide a way to identify novel gene expression patterns, new alternately spliced transcripts, and/or tissue-specific alleles for genes. Traditional approaches such as real-time reverse-transcriptase PCR (RT-PCR) or microarrays are limited to known genes and smaller datasets. Whole transcriptome sequencing provides the big picture, but RNA-Seq can also be used on subsets of genes. The addition of either a predesigned panel such as the xGen Whole Exome Hyb Capture Panel, or the xGen Custom Hyb Panel separates the relevant transcripts from the rest. Using a hybridization capture enrichment strategy increases flexibility in experimental designs.
RNA-Seq starts with library preparation, and IDT offers a variety of tools for studying gene expression and genotyping, including:
For low inputs (~100 pg mRNA or ~10 ng total RNA), deepen your rare and novel transcript discovery with the xGen Broad-Range RNA Library Prep Kit. Benefit from high mapping rates, more genes and transcripts detected, and consistent function across a broad range of inputs.
For higher inputs (>5 ng mRNA or >100 ng total RNA), accelerate your gene expression profiling studies with the xGen RNA Library Kit. Benefit from an expedited ~3.5-hour workflow, consistent function across inputs, and reduced costs per library prep.