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Related Experiment Video

Updated: Jun 1, 2026

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons
11:40

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

Published on: November 14, 2018

Fluorogenic DNA sequencing in PDMS microreactors.

Peter A Sims1, William J Greenleaf, Haifeng Duan

  • 1Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.

Nature Methods
|June 14, 2011
PubMed
Summary
This summary is machine-generated.

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We developed a novel sequencing-by-synthesis method using fluorogenic nucleotides and microreactors for rapid DNA analysis. This approach achieves high accuracy and sensitivity, offering a simplified alternative to existing DNA sequencing technologies.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genomics

Background:

  • DNA sequencing is crucial for biological research and diagnostics.
  • Existing methods can be complex, costly, or time-consuming.
  • Need for sensitive, accurate, and easily parallelizable sequencing techniques.

Purpose of the Study:

  • To develop a novel multiplex sequencing-by-synthesis method.
  • To combine terminal phosphate-labeled fluorogenic nucleotides (TPLFNs) with polydimethylsiloxane (PDMS) microreactors.
  • To achieve high-throughput, accurate, and sensitive DNA sequencing.

Main Methods:

  • Developed a method using TPLFNs and resealable PDMS microreactors.
  • Immobilized DNA templates and sequentially introduced TPLFNs.

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Published on: March 13, 2016

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules
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A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules

Published on: June 20, 2020

Related Experiment Videos

Last Updated: Jun 1, 2026

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons
11:40

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

Published on: November 14, 2018

Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles
11:13

Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles

Published on: March 13, 2016

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules
10:45

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules

Published on: June 20, 2020

  • Detected fluorophore generation via DNA polymerase extension within microreactors.
  • Utilized phosphatase for TPLFN activation and fluorescence generation.

    • Main Results:

    • Demonstrated 30 base reads with approximately 99% raw accuracy.
    • Achieved cycle times of less than 10 minutes.
    • Enabled simultaneous real-time monitoring of all microreactors, not required.

    Conclusions:

    • The 'fluorogenic pyrosequencing' method offers benefits of pyrosequencing, including rapid turnaround and native DNA generation.
    • Achieved high detection sensitivity and simplicity of parallelization.
    • Provides a sensitive, accurate, and simplified DNA sequencing approach.