The massively parallel sequencing technology known as next-generation sequencing (NGS) has revolutionized the biological sciences. With its ultra-high throughput, scalability, and speed, NGS enables researchers to perform a wide variety of applications and study biological systems at a level never before possible.
Today's complex genomic research questions demand a depth of information beyond the capacity of traditional DNA sequencing technologies. Next-generation sequencing has filled that gap and become an everyday research tool to address these questions.
NGS 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 researchers to:
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.
NGS makes sequence-based gene expression analysis a “digital” alternative to analog techniques. It lets you quantify RNA expression with the breadth of a microarray and the resolution of qPCR.
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.
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 specific 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.
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Illumina sequencing 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 sequence data that's richer and more complete than is imaginable with Sanger sequencing. 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.
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 Introduction
Recent Illumina next-generation sequencing technology breakthroughs include:
*This specification is based on a dual flow cell run of S4 flow cells which have not been released; therefore, performance metrics are subject to change.
The resources below offer valuable guidance to researchers who are considering purchasing an NGS system.