How does agarose gel electrophoresis separate dna

Abstract Agarose gel electrophoresis is the most effective way of separating DNA fragments of varying sizes ranging from bp to 25 kb 1. Publication types Video-Audio Media. Substances DNA. This technique might be used to separate proteins that have the same molecular weight but different charges, or when size is not important e. These days, charge IEF and size SDS-PAGE separation are often employed together in two-dimensional electrophoresis, where charge separation is first used, and then these separated proteins are separated on the basis on size.

This is a very effective method for identifying a particular protein from a tissue that may contain thousands of proteins and where there may only be small differences between control and treated samples e. Add to collection. Go to full glossary Add 0 items to collection. Download 0 items. Twitter Pinterest Facebook Instagram. Email Us. See our newsletters here.

Typically, a DNA molecule is digested with restriction enzymes , and the agarose gel electrophoresis is used as a diagnostic tool to visualize the fragments.

An electric current is used to move the DNA molecules across an agarose gel, which is a polysaccharide matrix that functions as a sort of sieve. The matrix helps "catch" the molecules as they are transported by the electric current.

This technique has lots of applications. Generally speaking you can analyze DNA fragments that result from an enzyme digestion of a larger piece of DNA to visualize the fragments and determine the sizes of the fragments. In addition to its usefulness in research techniques, agarose gel electrophoresis is a common forensic technique and is used in DNA fingerprinting.

Ethidium bromide is an intercalating dye, which means it inserts itself between the bases that are stacked in the center of the DNA helix. One ethidium bromide molecule binds to one base. As each dye molecule binds to the bases the helix is unwound to accommodate the strain from the dye. Ethidium bromide can easily get into your cells. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Lesley A. Mitchenall, Michael M. Qiagen Ltd. You can also search for this author in PubMed Google Scholar. Correspondence to Anthony Maxwell. Reprints and Permissions. Mitchenall, L. A rapid high-resolution method for resolving DNA topoisomers. BMC Res Notes 11, 37 Download citation. Received : 06 September Accepted : 09 January Published : 16 January Anyone you share the following link with will be able to read this content:.

Sorry, a shareable link is not currently available for this article. Provided by the Springer Nature SharedIt content-sharing initiative. Skip to main content. Search all BMC articles Search. Download PDF.

Mitchenall 1 , Rachel E. Hipkin 2 nAff3 , Michael M. Piperakis 1 nAff4 , Nicolas P. Abstract Objective Agarose gel electrophoresis has been the mainstay technique for the analysis of DNA samples of moderate size. Full size image. Results and discussion To assess the ability of the QIAxcel system to resolve different topological forms of closed-circular DNA, we generated a set of plasmid pBR samples bp with a wide distribution of linking numbers.

Limitations There are both advantages and potential disadvantages in using the QIAxcel system. References 1. PubMed Google Scholar 4. Google Scholar 7. CAS Google Scholar Chapter Google Scholar Availability of data and materials All data is given in the main body of the manuscript; materials are available from the authors. Consent to publish Not applicable. Ethical approval and consent to participate Not applicable.

Author information Author notes Rachel E. Burton Authors Lesley A. Mitchenall View author publications. View author publications. About this article.