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Total RNA Sequencing

Introduction to Total RNA Sequencing

Whole-transcriptome analysis with total RNA sequencing (RNA-Seq) detects coding plus multiple forms of noncoding RNA. Total RNA-Seq can accurately measure gene and transcript abundance, and identify known and novel features of the transcriptome.

Total RNA-Seq provides optimal coverage in normal or low-quality samples. Species-specific ribosomal RNA probes can efficiently remove abundant RNA species. This leaves both fragmented and intact transcripts of interest for library capture.

Advantages of Total RNA Sequencing

Advantages of Total RNA Sequencing

Total RNA-Seq analyzes both coding and multiple forms of noncoding RNA for a comprehensive view of the transcriptome.

  • Captures both known and novel features
  • Allows researchers to identify biomarkers across the broadest range of transcripts
  • Enables a more comprehensive understanding of phenotypes of interest
  • Allows profiling of the whole transcriptome across a wide dynamic range
Transitioning from Whole-Genome Arrays to Total RNA Sequencing

Transitioning from Whole-Genome Arrays to Total RNA-Seq

Dr. Avrum Spira, a pulmonary and critical care physician at Boston University Medical Center, started a lab with the goal of analyzing samples with whole-genome gene expression arrays, to study potential biomarkers for pulmonary disorders. Learn how Spira’s lab uses total RNA-Seq to uncover novel and rare transcripts that may not be detectable on expression arrays.
Read Case Study.

Deciphering the Role of Long Non-Coding RNA in Cancer

Researchers at Biogazelle are using RNA-Seq to reveal how lncRNAs can be specific to cell types. They discovered that silencing certain lncRNAs can help destroy cancer cells.

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Total RNA Sequencing Workflow

Illumina sequencing by synthesis (SBS) chemistry is the most widely adopted next-generation sequencing (NGS) technology, producing approximately 90% of global sequencing data.*

In addition to high-quality data, Illumina offers integrated whole-transcriptome sequencing workflows that simplify the entire process, from library preparation to data analysis and biological interpretation.

RNA Sequencing Considerations

Each RNA-Seq experiment type—whether it’s gene expression profiling, targeted RNA expression, or small RNA analysis—has unique requirements for read length and depth. This bulletin reviews experimental considerations and offers resources to help with study design.

Read Bulletin
RNA Sequencing Considerations
Using NGS to Study Rare Undiagnosed Genetic Disease

Using NGS to Study Rare Undiagnosed Genetic Disease

Whole-exome and transcriptome sequencing prove beneficial in uncovering mutations and pathways associated with rare disease

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Related Solutions

Cancer Research: Gene Expression Studies

Monitoring gene expression changes in both coding and noncoding RNA biomarkers with total RNA-Seq can help researchers understand which variants affect tumor classification and progression. Learn more about cancer RNA-Seq.

Agrigenomics: Deep Transcriptome Sequencing

Plants have large, repetitive genomes, which can make it difficult to sequence weakly expressed genes. Ultradeep RNA sequencing can facilitate analysis of these genes. Learn more about plant sequencing for agrigenomics.

Complex Disease Biomarker Identification

Total RNA-Seq allows complex disease researchers to study coding and multiple forms of noncoding RNA in a single analysis, providing visibility to a broad range of potential disease-associated biomarkers. Learn more about complex disease research.

RNA-Based Drug Response Biomarker Discovery

Find out how to utilize RNA-Seq to identify novel RNA-based drug response biomarkers. Access resources designed to help new users adopt this application. Learn more about RNA biomarker analysis for drug development.

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Additional Resources

RNA-Seq Data Analysis
RNA-Seq Data Analysis

User-friendly software tools simplify RNA-Seq data analysis for biologists, regardless of bioinformatics experience.

RNA-Seq of Low-Quality and FFPE Samples
RNA-Seq of Low-Quality and FFPE Samples

RNA-Seq of formalin-fixed, paraffin-embedded (FFPE) and other low-quality samples offers valuable insights for disease research.

RNA-Seq with the NextSeq 550 System
RNA-Seq with the NextSeq 550 System

A flexible, cost-effective solution for RNA sequencing provides a deeper understanding of biology.

Paired-End RNA-Seq
Paired-End RNA-Seq

All Illumina sequencing systems are capable of paired-end sequencing, which facilitates detection of novel RNA transcripts, gene fusions, and more.

*Data calculations on file. Illumina, Inc., 2015