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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.

Genome Annotation and Assembly

Genome Annotation and Assembly

The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.

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 microarray-based...

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
The...

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...

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Related Experiment Video

Updated: May 27, 2026

Novel Sequence Discovery by Subtractive Genomics

Novel Sequence Discovery by Subtractive Genomics

Published on: January 25, 2019

NCBI Reference Sequences (RefSeq): current status, new features and genome annotation policy.

Kim D Pruitt¹, Tatiana Tatusova, Garth R Brown

¹National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Building 38A, 8600 Rockville Pike, Bethesda, MD 20894, USA. pruitt@ncbi.nlm.nih.gov

Nucleic Acids Research

|November 29, 2011

Summary

This summary is machine-generated.

The National Center for Biotechnology Information (NCBI) Reference Sequence (RefSeq) database offers curated genomic, transcript, and protein records, reducing redundancy. It provides an updated view of sequence data, features, and cross-links for over 16,000 organisms.

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Area of Science:

Bioinformatics
Genomics
Molecular Biology

Background:

The NCBI RefSeq database is a vital resource for genomic, transcript, and protein sequences.
It aims to reduce redundancy found in public sequence archives.
RefSeq provides curated data for over 16,000 organisms, including prokaryotes, eukaryotes, and viruses.

Purpose of the Study:

To report on the recent growth and status of the RefSeq database.
To detail advancements in human RefSeq data curation and feature annotation.
To outline current policies for eukaryotic genome annotation using the NCBI annotation pipeline.

Main Methods:

Automated analyses and manual curation are employed to maintain the database.
Collaboration with other institutions aids in data selection and refinement.
The NCBI annotation pipeline is utilized for eukaryotic genome annotation.

Main Results:

RefSeq release 49 (September 2011) contains extensive genomic, protein, and RNA records.
Significant progress has been made in curating the human RefSeq dataset.
Enhanced feature annotation and updated policies for eukaryotic genome annotation are presented.

Conclusions:

The RefSeq database is a continuously growing and meticulously curated resource.
Ongoing efforts ensure an up-to-date representation of biological sequences and their features.
The NCBI annotation pipeline facilitates comprehensive eukaryotic genome annotation.