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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.

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Updated: May 29, 2026

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons

Published on: November 14, 2018

Droplet-based pyrosequencing using digital microfluidics.

Deborah J Boles¹, Jonathan L Benton, Germaine J Siew

¹Advanced Liquid Logic Incorporated, Research Triangle Park, North Carolina, United States.

Analytical Chemistry

|September 22, 2011

Summary

This summary is machine-generated.

Digital microfluidics enables droplet-based pyrosequencing, achieving 100% accuracy for over 60 bp. This low-cost platform demonstrates feasibility for DNA sequencing applications.

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Last Updated: May 29, 2026

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons

Published on: November 14, 2018

Digital Microfluidics for Automated Proteomic Processing

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Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

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Published on: September 24, 2015

Area of Science:

Biotechnology
Microfluidics
Genomics

Background:

Pyrosequencing is a widely used DNA sequencing method.
Digital microfluidics offers precise control over small fluid volumes.
Integrating these technologies could enhance sequencing efficiency.

Purpose of the Study:

To demonstrate the feasibility of pyrosequencing chemistry within droplets using electrowetting-based digital microfluidics.
To optimize reagents and protocols for enhanced signal and efficiency.
To assess the performance of this integrated platform for DNA sequencing.

Main Methods:

Utilized an electrowetting-based digital microfluidic platform with an electrode array.
Controlled droplet formation, transport, merging, mixing, and splitting for pyrosequencing.
Implemented a three-enzyme pyrosequencing protocol with magnetic bead-anchored DNA templates.
Optimized reagents and protocols for signal, linearity, and wash efficiency.

Main Results:

Successfully sequenced a portion of a Candida parapsilosis template using both de novo and resequencing protocols.
Achieved over 60 bp of sequence with 100% accuracy in the resequencing protocol.
Observed excellent linearity for homopolymers in the sequenced DNA.
Demonstrated efficient washing of magnetic bead-bound DNA templates.

Conclusions:

Electrowetting-based digital microfluidics is a feasible platform for droplet-based pyrosequencing.
The developed protocol shows high accuracy and linearity for short DNA sequences.
Future improvements in microfluidic design could lead to longer reads and higher throughput, enabling "sample-to-sequence" capability on a low-cost platform.