xGen™ NGS RNA-Seq Library Preparation

Overview

RNA sequencing (RNA-Seq) is a powerful tool with a wide range 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
• mRNA-focused gene expression analysis

RNA-Seq has enabled transcriptomics, the study of the full complement of RNA within a sample. Sequencing RNA provides a snapshot of which genes are expressed in a particular tissue, cell type, or even a single cell. Changes in the organism or cellular microenvironment can turn genes on or off, and studying these changes in gene expression helps researchers better understand genomics, cellular regulation, and organism function.

RNA-Seq strategies use NGS technology to study the transcriptome and identify novel gene expression patterns, alternatively spliced transcripts, tissue-specific alleles, and other RNA-level variations. Traditional approaches such as real-time reverse-transcriptase PCR (RT-PCR) or microarrays are limited to known targets and smaller datasets. Whole transcriptome sequencing provides a broader view, while targeted RNA sequencing can focus analysis on specific transcripts, genes, or pathways. IDT xGen hybridization capture reagents, including predesigned and custom panels, works with compatible RNA library prep workflows to help enrich relevant transcripts and support flexible experimental designs.

RNA-Seq starts with library preparation, and IDT offers a connected transcriptomics workflow designed to help researchers move from RNA to sequencing-ready libraries with speed, flexibility, and confidence. IDT solutions support whole-transcriptome profiling, mRNA-focused gene expression analysis, and targeted transcript analysis when hybridization capture enrichment is needed.

For low, variable, or precious RNA inputs, the xGen Broad-Range RNA Library Prep Kit enables rare and novel transcript discovery across a broad range of inputs. Benefit from high mapping rates, detection of more genes and transcripts, and consistent performance across challenging sample types and input amounts.

For higher-input, routine RNA-Seq workflows, the xGen RNA Library Kit helps accelerate gene expression profiling studies with an expedited ~3.5-hour workflow, consistent performance across inputs, and reduced cost per library prep.

For mRNA-focused gene expression analysis from total RNA, the xGen mRNA Enrichment Module enriches poly(A)-tailed mRNA before library preparation, helping direct sequencing reads toward mRNA transcripts and reduce reads spent on abundant rRNA. The module is compatible with both xGen RNA Library Prep and xGen Broad-Range RNA Library Prep, giving researchers added flexibility to pair mRNA enrichment with the library prep workflow that best fits their sample input, study design, and throughput needs.