Next-Generation Sequencing for Beginners
We'll guide you through the basics of NGS, with tutorials and tips for planning your first experiment.
Get StartedWhat is NGS?
Next-generation sequencing (NGS) is a massively parallel sequencing technology that offers ultra-high throughput, scalability, and speed. The technology is used to determine the order of nucleotides in entire genomes or targeted regions of DNA or RNA. NGS has revolutionized the biological sciences, allowing labs to perform a wide variety of applications and study biological systems at a level never before possible.
Today's complex genomics questions demand a depth of information beyond the capacity of traditional DNA sequencing technologies. NGS has filled that gap and become an everyday tool to address these questions.
Applications of NGS
Next-generation sequencing technology has fundamentally changed the kinds of questions scientists can ask and answer. Innovative sample preparation and data analysis options enable a broad range of applications. For example, NGS allows labs to:
- Rapidly sequence whole genomes
- Deeply sequence target regions
- Utilize RNA sequencing (RNA-Seq) to discover novel RNA variants and splice sites, or quantify mRNAs for gene expression analysis
- Analyze epigenetic factors such as genome-wide DNA methylation and DNA-protein interactions
- Sequence cancer samples to study rare somatic variants, tumor subclones, and more
- Study the human microbiome
- Identify novel pathogens
How Does Illumina NGS Work?
Illumina NGS technology utilizes a fundamentally different approach from the classic Sanger chain-termination method. It leverages sequencing by synthesis (SBS) technology – tracking the addition of labeled nucleotides as the DNA chain is copied – in a massively parallel fashion.
Next-generation sequencing generates masses of DNA sequencing data, and is both less expensive and less time-consuming than traditional Sanger sequencing.2 Illumina sequencing systems can deliver data output ranging from 300 kilobases up to multiple terabases in a single run, depending on instrument type and configuration.
In-Depth NGS Introduction
This detailed overview of Illumina sequencing describes the evolution of genomic science, major advances in sequencing technology, key methods, the basics of Illumina sequencing chemistry, and more.
Read IntroductionNGS Workflow
Perform next-generation sequencing workflow using three basic steps: library preparation, sequencing, and data analysis.
Library Preparation
Library Preparation Kits
Novel, easy-to-use, and fast solutions for both DNA and RNA library preparation.
NGS Library Prep Education
Learn how to prepare libraries for NGS using technologies enabled by continuous innovation.
Sequencing
Illumina NGS Platforms
Explore benchtop and production-scale instruments designed to help you choose the right platform for your needs.
Data Analysis
NGS Data Analysis Tools
User-friendly and intuitive Illumina tools simplify sequencing data analysis, allowing you to focus on research and spend less time configuring workflows.
Methods Guide
Access the information you need—from BeadChips to library preparation for genome, transcriptome, or epigenome studies to sequencer selection, analysis, and support—all in one place. Select the best tools for your lab with our comprehensive guide designed specifically for research applications.
Access Guide
Key Benefits of NGS
Accessible Whole-Genome Sequencing
Using capillary electrophoresis-based Sanger sequencing, the Human Genome Project took over 10 years and cost nearly $3 billion.
Next-generation sequencing, in contrast, makes large-scale whole-genome sequencing (WGS) accessible and practical for the average researcher. It enables scientists to analyze the entire human genome in a single sequencing experiment, or sequence thousands to tens of thousands of genomes in one year.
Broad Dynamic Range for Expression Profiling
NGS-based RNA-Seq is a powerful method that enables researchers to break through the inefficiency and expense of legacy technologies such as microarrays. Microarray gene expression measurement is limited by noise at the low end and signal saturation at the high end.
In contrast, next-gen sequencing quantifies discrete, digital sequencing read counts, offering a broader dynamic range.1,2,3
Tunable Resolution for Targeted NGS
Targeted sequencing allows you to sequence a subset of genes or specific genomic regions of interest, efficiently and cost-effectively focusing the power of NGS. NGS is highly scalable, allowing you to tune the level of resolution to meet experimental needs. Choose whether to do a shallow scan across multiple samples, or sequence at greater depth with fewer samples to find rare variants in a given region.
Learn more about:
Advances in NGS Technology
Recent Illumina next-generation sequencing technology breakthroughs include:
- Semiconductor sequencing: The iSeq 100 System combines a complementary metal-oxide semiconductor (CMOS) chip with one-channel SBS to deliver high-accuracy data in a compact system.
- Patterned flow cell technology: This option offers an exceptional level of throughput for diverse sequencing applications.
- 75 breakthrough innovations: The NextSeq 1000 and 2000 Systems offer flexibility for emerging applications, a simple workflow, and data analysis in as little as 2 hours.
- Up to 16 Tb: The NovaSeq X series provides extraordinary sequencing power to fuel data-intensive applications.
Precision Health
Personalized medicine programs can help match patients to treatments based on their genetic blueprints and improve survival rates, quality of life, and the cost of care.
Learn MoreHow Scientists Use NGS
See how researchers in different fields utilize next-generation sequencing to make breakthrough discoveries.
Genetics of COVID-19 Susceptibility
This UK-wide study uses NGS to compare the genomes of severely and mildly ill COVID-19 patients, to help uncover genetic factors associated with susceptibility.
Exploring the Tumor Microenvironment
Scientists use single-cell NGS techniques to study cancer microenvironments, elucidate gene expression patterns, and gain insights into drug resistance and metastasis.
Cell-Free RNA as a Noninvasive Biomarker
This research highlights the broad potential of circulating cell-free RNA sequencing for biomarker discovery and noninvasive health monitoring.
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NGS for COVID-19
Next-generation sequencing is uniquely positioned in an infectious disease surveillance and outbreak model. Learn which NGS methods are recommended for detecting and characterizing SARS-CoV-2 and other respiratory pathogens, tracking transmission, studying co-infection, and investigating viral evolution.
Explore Coronavirus NGS Methods
Start Using NGS
System Buying Guidance
The resources below offer valuable guidance to scientists who are considering purchasing a next-generation sequencing system.
- View NGS platform comparison tables
- Use our interactive platform selection tool
- Read our NGS System Buyer's Guide to determine what factors to consider before making your purchase.
NGS Experimental Considerations
Learn about read length, coverage, quality scores, and other experimental considerations to help you plan your sequencing run.
Use our interactive tools to help you create a custom NGS protocol or select the right products and methods for your project.
Resources for High-Throughput NGS Labs
- High-Throughput Sequencing: Process more samples to improve statistical power. Cost-effectively run data-rich multiomics applications using the latest large-scale sequencers.
- Library Prep Automation: Explore automated liquid handling solutions designed to help labs prepare large quantities of NGS libraries.
- Lab Information Management Systems (LIMS): Automate workflows, integrate instruments, and manage samples.
- NGS Data Storage: Securely store vast quantities of NGS and other genomics data.
Interested in learning more about how to perform NGS? Sign up to have a sales rep contact you.
Webinars
Flexible Solutions Using NextSeq 1000 and 2000
Watch a recorded discussion on how the NextSeq 1000/2000 sequencers offer the flexibility and scalability to handle diverse applications and methods for efficient discoveries.
Redefining NGS in Cancer Research
Experts provide an overview of achievements and challenges of NGS in advancing cancer research, including a discussion on how an integrated multiomics approach can be used in future cancer diagnosis and treatment.
Unlocking the Power of Genomics for Surveillance of SARS-CoV-2 Variants
Learn about methods for SARS-CoV-2 surveillance, including requirements, workflow, and analysis.
Genomics News
Addressing the genomics education gap
With limited space in the curricula for genomics, residents and physicians are turning to other sources for training
Read Article
Using analytics to improve cancer diagnosis and therapy selection
Developing and automating workflows for analyzing, processing, and sharing genomic data among researchers and clinicians.
Read Interview
CGP validated in triple-negative breast cancer patients
Taiwanese study shows 500-gene panel detects actionable mutations and other critical biomarkers to open doors to advanced treatments
Read ArticleRelated Solutions
NGS Library Preparation
Fast, simple NGS library prep and enrichment workflows from Illumina to prepare your samples for sequencing.
Sequencing Services
Access fast, reliable next-generation sequencing services that provide high-quality data and offer extensive scientific expertise.
Illumina NGS & Microarray Training
Work with expert Illumina instructors and get hands-on training. We also offer online courses, webinars, videos, and podcasts.