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Two-dimensional Gel Electrophoresis

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...

Electrophoresis: Overview

Electrophoresis: Overview

Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...

DNA Agarose Gel Electrophoresis

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
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SDS-PAGE

SDS-PAGE

Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
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Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
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Related Experiment Video

Updated: Jul 15, 2026

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Published on: February 24, 2011

Pulsed-field gel electrophoresis.

Jill Herschleb¹, Gene Ananiev, David C Schwartz

¹Department of Chemistry, UW Biotechnology Center, University of Wisconsin-Madison, 425 Henry Mall, Madison, Wisconsin 53706, USA.

Nature Protocols

|April 5, 2007

Summary

Pulsed-field gel electrophoresis (PFGE) separates large DNA molecules up to 10 Mb, unlike standard methods. This technique is crucial for genomic analysis of microbes and mammalian cells.

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

Standard gel electrophoresis is limited to resolving DNA fragments up to approximately 50 kb.
Large DNA molecules exhibit unique behaviors within a gel matrix during electrophoresis.
Efficient separation of large DNA molecules is essential for advanced genomic studies.

Purpose of the Study:

To describe the protocol for pulsed-field gel electrophoresis (PFGE).
To detail the operation of specialized PFGE instrumentation.
To provide instructions for preparing intact chromosomal DNA for PFGE analysis.

Main Methods:

Pulsed-field gel electrophoresis (PFGE) utilizes precisely oriented electrical pulses to separate large DNA molecules (up to 10 Mb).
The method relies on the size-dependent perturbation of DNA molecule trajectories as they navigate a gel matrix.

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The protocol includes descriptions of custom-built PFGE instrumentation and DNA preparation techniques.

Main Results:

PFGE enables the separation of DNA molecules significantly larger than achievable with standard gel electrophoresis.
The described protocol facilitates the rapid genomic analysis of microbial and mammalian cells.
PFGE has spurred the development of large-insert cloning systems, including bacterial and yeast artificial chromosomes.

Conclusions:

PFGE is a powerful technique for the high-resolution separation of large DNA molecules.
The protocol provides a comprehensive guide for PFGE implementation and instrumentation.
This method is instrumental in advancing genomic research and molecular cloning strategies.