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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...

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...

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...

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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

Field guide to next-generation DNA sequencers.

Travis C Glenn¹

¹Department of Environmental Health Science and Georgia Genomics Facility, Environmental Health Science Building, University of Georgia, Athens, GA 30602, USA. travisg@uga.edu

Molecular Ecology Resources

|May 20, 2011

Summary

This summary is machine-generated.

Choosing the right DNA sequencing platform requires careful consideration of cost and speed trade-offs. Illumina and SOLiD offer the lowest cost per megabase, while Pacific Biosciences and Ion Torrent excel in speed and cost per sample.

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Area of Science:

Genomics
Molecular Biology
Bioinformatics

Background:

Rapidly increasing diversity of 2nd and 3rd generation DNA sequencing platforms.
Significant cost and performance trade-offs exist among available platforms.

Purpose of the Study:

To summarize and compare major characteristics of commercially available DNA sequencing platforms.
To guide informed decisions regarding platform selection for research and cost-efficiency.

Main Methods:

Comparative analysis of instrument costs, run times, and cost per megabase (Mb) and per sample.
Evaluation of multiplexing capabilities and library preparation considerations.

Main Results:

Illumina and SOLiD platforms demonstrate superior cost per Mb (≤ $0.10/Mb).
Pacific Biosciences and Ion Torrent platforms offer advantages in cost per non-multiplexed sample and instrument run time.
454 GS Junior and Illumina MiSeq are also notable for sample throughput and speed.

Conclusions:

Careful evaluation of desired platform characteristics is crucial before acquisition or use.
Informed selection can enable groundbreaking studies and prevent financial waste.
Platform selection depends on specific experimental needs, balancing cost, speed, and throughput.