Next-Generation Sequencing (NGS)

Next-generation sequencing for beginners

We'll guide you through the basics of NGS, with tutorials and tips for planning your first experiment.

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.

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

Explore Sequencing Methods & Uses

Illumina NGS technology utilizes a fundamentally different approach from the classic Sanger chain-termination method. It leverages sequencing by synthesis (SBS) chemistry – tracking the addition of labeled nucleotides as the DNA chain is copied – in a massively parallel fashion.

Next-generation sequencing technology generates masses of DNA sequencing data, and is both less expensive and less time-consuming than traditional Sanger sequencing.^1,2 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.

Learn more about SBS

Accessible whole-genome sequencing

Using capillary electrophoresis-based Sanger sequencing, the Human Genome Project took over 10 years and cost nearly 3ドル 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.

Learn more about WGS

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-generation sequencing quantifies discrete, digital sequencing read counts, offering a broader dynamic range.^3,4,5

Compare microarrays vs RNA-Seq

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:

• Targeted sequencing
• NGS vs Sanger sequencing
• NGS vs qPCR

Recent Illumina next-generation sequencing technology breakthroughs include:

• XLEAP-SBS chemistry: This innovation delivers increased speed and greater fidelity compared to standard Illumina sequencing by synthesis (SBS) chemistry.
  
• Up to 16 Tb: The NovaSeq X Series provides extraordinary sequencing power to fuel data-intensive applications.
• Breakthrough speed and simplicity: The MiSeq i100 Series offers benchtop sequencing run times as fast as four hours, with streamlined operations and intuitive onboard data analysis.
• Semiconductor sequencing: This technology combines a complementary metal-oxide semiconductor (CMOS) chip with SBS to deliver high-accuracy data in a compact instrument.
• Patterned flow cell technology: This advance provides an exceptional level of throughput for diverse sequencing applications.

Advancing drug discovery with microbiome studies

NGS-based whole-genome shotgun sequencing and transcriptomics provide researchers and pharmaceutical companies with data to refine drug discovery and development.

Read article

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.

Read article

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.

Read article

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.

Start Planning Experiments

• Sequencing methods: Explore a broad range of methods, from whole-genome sequencing to mRNA-Seq, exome sequencing, single-cell sequencing, and more.
• High-throughput sequencing: Process more samples to improve statistical power. Cost-effectively run data-rich 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.
• NGS data storage: Securely store vast quantities of NGS and other genomics data.

Genomics news

References

1. Bentley DR, Balasubramanian S, Swerdlow HP, et al. Accurate Whole Human Genome Sequencing using Reversible Terminator Chemistry. Nature. 2008; 456 (7218): 53–59.
2. Shendure J and Ji H. Next-generation DNA sequencing. Nat Biotechnol. 2008; 26(10): 1135-1145.
3. Wang Z, Gerstein M, Snyder M. RNA-Seq: a revolutionary tool for transcriptomics. Nat Rev Genet. 2009; 10: 57–63.
4. Wilhelm BT, Landry JR. RNA-Seq—quantitative measurement of expression through massively parallel RNA sequencing. Methods. 2009; 48: 249–57.
5. Zhao S, Fung-Leung WP, Bittner A, and Ngo K, Liu X. Comparison of RNA-Seq and microarray in transcriptome profiling of activated T cells. PLoS One. 2014; 16;9(1): e78644.