A highly sensitive method for measuring gene expression from single cells

Generating RNA-Seq libraries from single cell and ultra-low-input samples

Single-Cell and Ultra-Low-Input RNA-Seq

Complex biological systems are determined by the coordinated functions of individual cells. Conventional methods that provide bulk genome or transcriptome data are unable to reveal the cellular heterogeneity that drives this complexity. Single-cell sequencing is a next-generation sequencing (NGS) method that examines the genomes or transcriptomes of individual cells, providing a high-resolution view of cell-to-cell variation.

Highly sensitive ultra-low-input and single-cell RNA sequencing (RNA-Seq) methods enable researchers to explore the distinct biology of individual cells in complex tissues and understand cellular subpopulation responses to environmental cues. These assays enhance the study of cell function and heterogeneity in time-dependent processes such as differentiation, proliferation, and tumorigenesis.

Single-cell and ultra-low-input RNA-Seq are powerful tools for studying the transcriptome in an unbiased manner from minimal input.

  • Robust transcriptome analysis down to single-cell input levels for high-quality samples
  • Integrated protocol proceeds directly from whole cells and preserves sample integrity
  • High resolution analysis enables discovery of cellular differences usually masked by bulk sampling methods
Interested in valuable insights about the single-cell sequencing workflow?

Interested in valuable insights about the single-cell sequencing workflow?

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Featured Single-Cell NGS

 
Single-Cell Research Review
Single-Cell Research Review

These peer-reviewed publications demonstrate various single-cell sequencing applications and techniques using Illumina technology.

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The Singular Neuron
The Singular Neuron

James Eberwine explains how single-cell RNA sequencing can be used in vivo to understand how individual cells function in their microenvironment within a complex organism.

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Deciphering the Role of Long Non-Coding RNA in Cancer
Deciphering the Role of Long Non-Coding RNA in Cancer

Researchers are using RNA-Seq to reveal how lncRNAs could be used to identify, measure, and treat cancer.

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Illumina Tagment DNA TDE1 Enzyme and Buffer Kits

Use these components in ATAC-Seq experiments to analyze chromatin accessibility

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Illumina Tagment DNA TDE1 Enzyme and Buffer Kits

Single-cell sequencing methods can be distinguished by cell throughput. Low-throughput methods include mechanical manipulation or cell sorting/partitioning technologies and are able to process dozens to a few hundred cells per experiment.

Recent advances in microfluidic technologies have enabled high-throughput single cell profiling where researchers can examine hundreds to tens of thousands of cells per experiment in a cost-effective manner. Both the high- and low-throughput methods utilize Illumina sequencing by synthesis (SBS) chemistry, the most widely adopted NGS technology, which generates approximately 90% of sequencing data worldwide.*

The Singular Neuron

James Eberwine explains how single-cell RNA sequencing can be used in vivo to understand how individual cells function in their microenvironment within a complex organism.

View Video

The Illumina Bio-Rad Single-Cell RNA Sequencing Solution combines the highly innovative Bio-Rad Droplet Digital™ technology (ddSEQ™) with Illumina NGS library preparation, sequencing, and analysis technologies. Gain valuable insight into gene expression with this sensitive, scalable, and cost-effective high-throughput workflow.

Click on the below to view products for each workflow step.

ddSEQ™ Single-Cell Isolator

Scalable, flexible single-cell isolation to process tens of thousands of cells in a single day.

Illumina Bio-Rad® SureCell™ WTA 3′ Library Prep Kit for the ddSEQ™ System

Transcriptome profiling of hundreds to tens of thousands of single cells in a single experiment.

NextSeq 550 System

Flexible power and simplicity for whole-genome, exome, or transcriptome sequencing.

NovaSeq 6000 System

Scalable throughput and flexibility for virtually any genome, sequencing method, and scale of project.

Platform Comparison Tool

Compare sequencing platforms and identify the best system for your lab and applications.

Sequencing Reagents

Find reagents, flow cells, and buffers tailored to each Illumina sequencing system.

BaseSpace SureCell RNA Single-Cell App

Simple, yet powerful app enables streamlined primary and secondary analysis of single-cell RNA-Seq data.

SeqGeq™ Software by FlowJo, LLC

Explore cell sub-populations, identify differentially expressed genes, and generate publication-ready figures.

NovaSeq 6000 Sequencing System

The NovaSeq 6000 System combines benchtop sequencer ease of use with production-scale power in a single platform, with adjustable data output for maximum efficiency.

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NovaSeq 6000 Sequencing System
NovaSeq 6000 SP Reagent Kits

These reagent kits provide the flow cell and reagent consumables to support a single flow cell at 100, 300, or 500 cycles per NovaSeq run.

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NovaSeq 6000 S2 Reagent Kits

These reagent kits provide the flow cell and reagent consumables to support a single flow cell at 100, 200, or 300 cycles per NovaSeq run.

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Nextera XT DNA Library Preparation Kit

Prepare sequencing libraries for small genomes, PCR amplicons, plasmids, or cDNA in as little as 90 minutes, with a low DNA input requirement.

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The low-throughput method below is recommended for researchers who wish to process small numbers of cells for a particular study, such as dozens to a few hundred cells per experiment.

Click on the below to view products for each workflow step.

SMART-Seq® Ultra® Low Input RNA Kit

Synthesize full-length cDNA from 1–1000 whole cells or 10 pg–10 ng of high-quality total RNA. Developed by Clontech, sold by Takara, and optimized for Illumina sequencing.

Nextera XT DNA Library Preparation Kit

Optimal for preparing an Illumina RNA sequencing library from cDNA generated with the SMART-Seq Ultra Low Input RNA kit.

NextSeq 550 System

Flexible power and simplicity for whole-genome, exome, or transcriptome sequencing. 3-10 total RNA samples per run.

NovaSeq 6000 System

Scalable throughput and flexibility for virtually any genome, sequencing method, and scale of project.

Platform Comparison Tool

Compare sequencing platforms and identify the best system for your lab and applications.

Sequencing Reagents

Find reagents, flow cells, and buffers tailored to each Illumina sequencing system.

BaseSpace RNA Express App

Aligns RNA-Seq reads with the STAR aligner and assigns aligned reads to genes, followed by differential expression with DESeq2.

BaseSpace TopHat Alignment App

Maps reads, performs abundance estimations of reference genes and transcripts, calls variants, and offers optional fusion calling.

BaseSpace Cufflinks Assembly & Differential Expression (DE) App

Assembles novel transcripts and performs differential expression of novel and reference transcripts.

Integrative Genomics Viewer

A genome browser developed by the Broad Institute of MIT and Harvard that displays NGS data for complex variant analysis.

Genomatix Pathway System (GePS)

Associates single gene or list of genes with annotation data for pathways, diseases, tissues, and small molecules.

iPathway Guide

Differential gene expression, drug interaction, and disease analysis.

BaseSpace Sequence Hub

The Illumina genomics computing environment for NGS data analysis and management.

BaseSpace Correlation Engine

A growing library of curated genomic data to support researchers in identifying disease mechanisms, drug targets, and biomarkers.

NGS vs. qPCR

The key difference between next-generation sequencing and qPCR is discovery power. Compare the two technologies and learn how the wide dynamic range and high sensitivity of NGS can help you detect and quantify variation.

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NGS vs. qPCR
Marine Biology: Uncover Hidden Diversity
Marine Biology: Uncover Hidden Diversity

Researchers at Bigelow Laboratory for Ocean Sciences use single-cell RNA sequencing to study bacteria inhabiting the surface layers of the ocean. Learn more about single-cell RNA-Seq in marine research.

RNA-Seq for Cancer Research
Cancer Research: Study Individual Clonal Populations

Evaluating transcriptome profile differences within tumor regions can enhance researchers' understanding of relapse and metastasis. Learn more about cancer RNA-Seq.

Single-Cell Research Review
Single-Cell Research Review

See an overview of peer-reviewed publications using Illumina technology for single-cell sequencing. These publications demonstrate various single-cell sequencing applications and techniques. Read the review.

 
 
Analysis of Single-Cell RNA-Seq Identifies Cell-Cell Communication Associated with Tumor Characteristics

Researchers used single-cell RNA-Seq to characterize cell-to-cell communication via ligand-receptor interactions across cell types in a tumor microenvironment.

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Human haematopoietic stem cell lineage commitment is a continuous process

Researchers used single-cell RNA-Seq to demonstrate that hematopoietic stem cell lineage commitment is a gradual process without differentiation into discrete progenitors.

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Aging increases cell-to-cell transcriptional variability upon immune stimulation

AResearchers used single-cell RNA-Seq to explore the effects of aging on the immune system, observing that age-related cell-to-cell transcriptional variability is a hallmark of aging.

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The Illumina Bio-Rad Single Cell Sequencing Solution
The Illumina Bio-Rad Single Cell Sequencing Solution

Profile transcriptomes of hundreds to tens of thousands of single cells with this user-friendly solution for gene expression studies.

Watch Video
Single-Cell RNA Sequencing of Peripheral Blood Mononuclear Cells
Single-Cell RNA Sequencing of Peripheral Blood Mononuclear Cells

Gain insight into how individual cells contribute to the function of a complex tissue such as peripheral blood.

Read App Note
RNA-Seq Data Analysis
RNA-Seq Data Analysis

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

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Single-Cell RNA-Seq and Stem Cell Research
Stem Cell Research with Single-Cell RNA-Seq

See how single-cell sequencing is transforming our view of cellular development in stem cells.

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Mapping Neural Diversity with Single-Cell mRNA-Seq
Mapping Neural Diversity with Single-Cell mRNA-Seq

The ability to analyze gene expression signatures from individual cells is transforming the way neurons are classified.

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Single-Cell RNA-Seq, the Internet, and Memory
Single-Cell RNA-Seq, the Internet, and Memory

TGen researchers use single-cell RNA sequencing to understand how individual neurons are involved in memory formation.

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Single-Cell Sequencing on the NovaSeq 6000 System
Single-Cell Sequencing on the NovaSeq 6000 System

This scalable, robust, single-cell NGS methodology enables routine transcriptome profiling at single-cell resolution.

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Single-Cell Sequencing: Needle in a Haystack
Single-Cell Sequencing: Needle in a Haystack

Single-cell analysis studies are on the rise with applications in a myriad of fields.

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