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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
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Genomics

Genomics

Genomics is the science of genomes: it is the study of all the genetic material of an organism. In humans, the genome consists of information carried in 23 pairs of chromosomes in the nucleus, as well as mitochondrial DNA. In genomics, both coding and non-coding DNA is sequenced and analyzed. Genomics allows a better understanding of all living things, their evolution, and their diversity. It has a myriad of uses: for example, to build phylogenetic trees, to improve productivity and...

Sanger Sequencing

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

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.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...

Maxam-Gilbert Sequencing

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
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Evolutionary Relationships through Genome Comparisons

Evolutionary Relationships through Genome Comparisons

Genome comparison is one of the excellent ways to interpret the evolutionary relationships between organisms. The basic principle of genome comparison is that if two species share a common feature, it is likely encoded by the DNA sequence conserved between both species. The advent of genome sequencing technologies in the late 20th century enabled scientists to understand the concept of conservation of domains between species and helped them to deduce evolutionary relationships across diverse...

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Related Experiment Video

Updated: Jun 25, 2025

Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease

Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease

Published on: April 4, 2018

The Third-Generation Sequencing Challenge: Novel Insights for the Omic Sciences.

Carmela Scarano^1,2, Iolanda Veneruso^1,2, Rosa Redenta De Simone^1,2

¹Department of Molecular Medicine and Medical Biotechnologies, Federico II University, Via Sergio Pansini 5, 80131 Napoli, Italy.

|May 24, 2024

Summary

Third-generation sequencing (TGS) offers longer read lengths and improved accuracy, overcoming limitations of next-generation sequencing (NGS). TGS shows great potential for genomic analysis and diagnostics.

Keywords:

Oxford Nanopore Technologies PacBio RNA sequencing epigenetics genome sequencing metagenomics third-generation sequencing

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Last Updated: Jun 25, 2025

Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease

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Published on: April 4, 2018

Leveraging CyVerse Resources for De Novo Comparative Transcriptomics of Underserved Non-model Organisms

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Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Published on: June 3, 2019

Area of Science:

Genomics
Molecular Biology
Bioinformatics

Background:

Next-generation sequencing (NGS) advanced genomic understanding but has limitations like short read length and PCR dependence.
Third-generation sequencing (TGS) technologies were developed to address NGS constraints, offering significant improvements.

Purpose of the Study:

To highlight the advancements and potential applications of TGS in genomics.
To discuss how TGS overcomes the limitations of previous sequencing methods.

Main Methods:

Review of TGS technology evolution, focusing on improvements in read length, accuracy, and cost.
Exploration of TGS applications across various genomic analyses.

Main Results:

TGS has progressively enhanced read length, base-calling accuracy, and reduced costs per base.
TGS demonstrates significant potential in analyzing complex genomic regions, structural variations, RNA expression, DNA methylation, and metagenomics.

Conclusions:

TGS is a powerful tool with expanding applications in research and potential for routine diagnostics.
Standardization of protocols and user-friendly data analysis pipelines are crucial for widespread TGS adoption.