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Two-dimensional Gel Electrophoresis01:22

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: Overview01:20

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 Electrophoresis02:35

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.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
SDS-PAGE01:27

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.
A variation of gel electrophoresis, termed  polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Capillary Electrophoresis: Instrumentation01:20

Capillary Electrophoresis: Instrumentation

Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...

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Updated: Jun 3, 2026

Detection of Bacteria Using Fluorogenic DNAzymes
13:20

Detection of Bacteria Using Fluorogenic DNAzymes

Published on: May 28, 2012

Pulsed-field gel electrophoresis.

M E Kaufmann1

  • 1Laboratory of Hospital Infection, Central Public Health Laboratory, London, UK.

Methods in Molecular Medicine
|March 11, 2011
PubMed
Summary

Pulsed-field gel electrophoresis (PFGE) enables the separation of large DNA fragments up to 10 megabases. This technique uses alternating electric fields to reorient DNA, allowing size-based separation unlike conventional methods.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Conventional agarose gel electrophoresis has limitations in resolving large DNA fragments (>20 kb).
  • Pulsed-field gel electrophoresis (PFGE) was developed to overcome these limitations.

Purpose of the Study:

  • To describe the principles and methodology of Pulsed-field gel electrophoresis (PFGE).
  • To highlight the advantages of PFGE for separating large DNA fragments.

Main Methods:

  • Preparation of unsheared DNA.
  • Digestion with rare-cutting restriction endonucleases.
  • Separation of DNA fragments using alternating electric fields in PFGE.

Main Results:

  • PFGE can resolve DNA fragments up to 10 megabases (Mb), significantly larger than conventional methods.

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Agarose Gel Electrophoresis for the Separation of DNA Fragments
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Agarose Gel Electrophoresis for the Separation of DNA Fragments

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Denaturing Gradient Gel Electrophoresis (DGGE)
10:52

Denaturing Gradient Gel Electrophoresis (DGGE)

Published on: February 25, 2007

Related Experiment Videos

Last Updated: Jun 3, 2026

Detection of Bacteria Using Fluorogenic DNAzymes
13:20

Detection of Bacteria Using Fluorogenic DNAzymes

Published on: May 28, 2012

Agarose Gel Electrophoresis for the Separation of DNA Fragments
07:10

Agarose Gel Electrophoresis for the Separation of DNA Fragments

Published on: April 20, 2012

Denaturing Gradient Gel Electrophoresis (DGGE)
10:52

Denaturing Gradient Gel Electrophoresis (DGGE)

Published on: February 25, 2007

  • Separation is based on the time DNA fragments take to reorient to changing electric fields, proportional to molecular weight.
  • Conclusions:

    • PFGE is a powerful technique for the separation and analysis of large DNA molecules.
    • The method is crucial for applications requiring the resolution of genome-sized DNA fragments.