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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.

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...

Multi-species Conserved Sequences

Multi-species Conserved Sequences

Next-generation sequencing technologies have created large genomic databases of a variety of animals and plants. Ever since the human genome project was completed, scientists studied the genome of primates, mammals, and other phylogenetically distant living beings. Such large-scale studies have provided new insights into the evolutionary relationship between organisms.
Although the genome of each species varies greatly from each other, a few sequences are highly conserved. Such conserved DNA...

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.
Challenges of the Maxam-Gilbert Method
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Cis-regulatory Sequences

Cis-regulatory Sequences

Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...

Cis-regulatory Sequences

Cis-regulatory Sequences

Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...

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Related Experiment Video

Updated: Jun 28, 2026

Novel Sequence Discovery by Subtractive Genomics

Novel Sequence Discovery by Subtractive Genomics

Published on: January 25, 2019

NCBI Reference Sequences: current status, policy and new initiatives.

Kim D Pruitt¹, Tatiana Tatusova, William Klimke

¹National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Rm 4As.47B, 45 Center Drive, Bethesda, MD, USA. pruitt@ncbi.nlm.nih.gov

Nucleic Acids Research

|October 18, 2008

Summary

The National Center for Biotechnology Information (NCBI) Reference Sequence (RefSeq) database has grown significantly, now including over 5.5 million proteins. A new initiative, RefSeqGene, will support reporting variation data.

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Area of Science:

Bioinformatics
Genomics
Molecular Biology

Background:

The NCBI Reference Sequence (RefSeq) database is a vital, curated repository of genomic, transcript, and protein sequences.
It encompasses over 5300 organisms and 5.5 million protein records, serving as a comprehensive biological data resource.

Purpose of the Study:

To report on the recent expansion and enhancements of the RefSeq database.
To detail updates in feature annotations and record types for eukaryotic species.
To introduce RefSeqGene for stable genomic coordinate systems and variation data reporting.

Main Methods:

Database curation and integration of information from multiple sources.
Application of feature annotation through curation, collaboration, propagation, and computation.
Development of new initiatives like RefSeqGene for specific data management needs.

Main Results:

Significant growth in the RefSeq database, including an expanded collection of protein records.
Recent modifications to feature annotations and record types, particularly for vertebrate species.
Introduction of RefSeqGene to standardize reporting of variation data.

Conclusions:

The RefSeq database continues to expand and evolve, providing a robust and up-to-date resource for biological research.
The RefSeqGene initiative promises to improve the management and accessibility of genomic variation data.