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Related Concept Videos

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...
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...
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...

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Related Experiment Video

Updated: Jul 5, 2026

Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)
08:44

Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)

Published on: October 29, 2009

Denaturing polyacrylamide gel electrophoresis.

L M Albright1, B E Slatko

  • 1Allison Park, Pennsylvania, USA.

Current Protocols in Human Genetics
|April 23, 2008
PubMed
Summary
This summary is machine-generated.

High-concentration urea polyacrylamide gels effectively separate short DNA and RNA fragments differing by a single nucleotide. This protocol details preparing and running these gels for various molecular analyses, including DNA sequencing and mutation detection.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Polyacrylamide gels with high urea concentrations are crucial for high-resolution separation of nucleic acid fragments.
  • These gels are widely applied in DNA sequencing, mutation detection, and genotyping techniques.

Purpose of the Study:

  • To provide a detailed protocol for preparing and running polyacrylamide-urea gels.
  • To enable precise separation of short single-stranded DNA or RNA fragments.

Main Methods:

  • Utilizing polyacrylamide gels with 7 M urea and 4%-8% acrylamide concentrations.
  • Describing the pouring, running, and processing of 40-cm long, 0.4-mm thick gels.

Main Results:

  • Demonstrated capability to resolve single-nucleotide length differences in DNA/RNA fragments (<500 nucleotides).
  • Established a reliable method for preparing gels suitable for various molecular biology applications.

Conclusions:

  • The described protocol facilitates accurate analysis of DNA and RNA at single-nucleotide resolution.
  • These gels are essential tools for applications such as DNA sequencing, PCR amplification of simple sequence length polymorphisms (SSLPs), ligase chain reaction (LCR) genotyping, and mutation analysis.