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Next-generation Sequencing

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.

RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
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Sanger Sequencing

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Related Experiment Video

Updated: Jun 8, 2026

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Published on: January 7, 2020

Multiplex parallel pair-end-ditag sequencing approaches in system biology.

Yijun Ruan¹, Chia-Lin Wei¹

¹Genome Technology & Biology Group, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672.

Wiley Interdisciplinary Reviews. Systems Biology and Medicine

|September 14, 2010

Summary

Pair end ditag (PET) sequencing overcomes short read limitations of next-generation sequencing. This technology enables comprehensive genome analysis, aiding in personalized medicine strategies.

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Area of Science:

Genomics and Bioinformatics
Molecular Biology
Systems Biology

Background:

Understanding the human genome requires elucidating complex regulatory networks, chromatin states, nuclear architectures, and variations.
Next-generation sequencing (NGS) technologies offer high throughput but are limited by short read lengths for comprehensive analysis.
Short read lengths hinder detailed studies of genome organization and regulatory elements.

Purpose of the Study:

To introduce and validate the pair end ditag (PET) sequencing strategy as a solution to NGS limitations.
To demonstrate PET sequencing's capability in expanding information content and linear coverage of the genome.
To highlight PET sequencing's potential in advancing system biology, genome organization, and personalized medicine.

Main Methods:

Sequencing of paired end signatures from DNA fragments.
Mapping of sequenced fragments to a reference genome.
Utilizing the information from paired ends to define DNA boundaries and relationships.

Main Results:

PET sequencing accurately demarcates target DNA boundaries and their genomic locations.
The strategy enables the discovery of unconventional gene products, genome rearrangements, and chromatin interactions.
PET strategy, when coupled with high-throughput sequencing, offers a powerful approach for genome-wide analyses.

Conclusions:

PET sequencing overcomes the limitations of short-read NGS, providing extended linear coverage and richer information content.
This technology revolutionizes the study of regulatory networks, genome organization, and variations.
PET sequencing holds significant potential for the development of personalized medicine strategies.