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Sanger Sequencing

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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...

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An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing

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Long-Read, High-Resolution Sanger Sequencing by Micelle-Tagging Electrophoresis.

Randall Gamble¹, H Michael Wenz², Bashar Mullah²

¹Department of Chemical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.

Electrophoresis

|October 22, 2025

Summary

This study introduces micelle-tagging electrophoresis (MTE) for gel-free Sanger sequencing, extending read lengths up to 782 bases. This novel approach significantly improves DNA fragment separation efficiency in electrophoresis.

Keywords:

DNA electrophoresis Sanger sequencing capillary electrophoresis end‐labeled free‐solution electrophoresis

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Last Updated: Jan 14, 2026

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Published on: May 23, 2018

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Published on: March 15, 2019

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Published on: November 18, 2014

Area of Science:

Biochemistry
Analytical Chemistry
Molecular Biology

Background:

Electrophoretic separation is crucial for DNA sequencing.
Existing methods like end-labeled free-solution electrophoresis (ELFSE) have limitations in read length.
Micelle-tagging electrophoresis (MTE) offers a potential alternative for improved separation.

Purpose of the Study:

To demonstrate gel-free electrophoretic separation of Sanger sequencing fragments using nonionic wormlike micelles.
To significantly increase the read length achievable with MTE.
To improve upon existing free-solution electrophoresis methods.

Main Methods:

Utilized nonionic wormlike micelles as drag-tags in micelle-tagging electrophoresis (MTE).
Attached C18 alkane groups to primers for micelle binding.
Developed a two-parameter time-shifting procedure to align electropherograms.

Main Results:

Achieved electrophoretic separation of Sanger sequencing fragments up to 782 bases.
Demonstrated a 280-base increase over previous MTE methods.
Showed a nearly three-fold improvement compared to ELFSE methods.

Conclusions:

Micelle-tagging electrophoresis with nonionic wormlike micelles enables significantly longer DNA sequencing reads.
The uniform drag provided by micelles and the alignment procedure contribute to enhanced read length.
This gel-free method offers a substantial advancement in DNA sequencing technology.