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Nucleic Acid Gel Electrophoresis

DNA electrophoresis using an agarose gel

Nucleic acid gel electrophoresis is a molecular biology technique for the separation, identification, and purification of DNA and RNA fragments based on size and charge. In this technique, DNA and RNA molecules are separated by applying an electric field to move negatively charged nucleic acids through an agarose or polyacrylamide gel matrix.

The gel matrix acts as a sieve, allowing shorter molecules to run more quickly through the pores of the gel than longer molecules.

  • Agarose gels are more commonly used for DNA and RNA electrophoresis and can resolve DNA fragments ranging from 50 base pairs (50 bp) to 50,000 bp using standard electrophoretic techniques.
  • Polyacrylamide gels have a smaller range of separation and can resolve fragments of DNA less than ~500 bp with very high resolution.
  • Both agarose and polyacrylamide gels can also be used in electrophoretic mobility shift assays (EMSA) to characterize protein-nucleic acid interactions.

Following electrophoresis, DNA and RNA molecules can be visualized via the use of stains or UV light, extracted and purified from the gel, or transferred for blotting. These methods are common preparative techniques in cloning, PCR, mass spectrometry, next generation sequencing (NGS), and Northern and Southern blot applications and workflows.

Related Technical Articles

Related Protocols

  • An introduction to both Northern and Southern blotting, popular methods for the transfer of macromolecules to membranous support. This article also offers a Southern blot protocol and a northern blot protocol.
  • The cloning process requires the ligation of linear DNA into a cloning vector. This ability to join fragments of DNA through recombinant technology is essential for many basic experiments in biotechnology.
  • A quick, clean and easy to use alternative to using a scalpel or razor blade for the extraction of DNA and RNA bands from agarose gels following gel electrophoresis.
  • Removal of Ethidium Bromide from DNA by Extraction with Organic Solvents Protocol from Cold Spring Harbor Laboratory Press.
  • The GenElute Gel Extraction kit is designed for the rapid purification of 50 bp to 10 kb linear DNA fragments and plasmids from standard or low-melting agarose gels.
  • This product is suitable for DNA precipitation from low salt aqueous buffers. It has been tested for the absence of proteases and nucleases.
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