Electrophoresis and Blotting of DNA

Sandeep Tamber, University of British Columbia, Vancouver, Canada
Robert EW Hancock, University of British Columbia, Vancouver, Canada

Published online: March 2004

DOI: 10.1038/npg.els.0003746

DNA electrophoresis and blotting are techniques commonly used to visualize DNA. Electrophoresis uses gels to separate DNA molecules on the basis of size; whereas blotting allows researchers to identify specific DNA fragments on the basis of their sequences.

Keywords: DNA detection; DNA analysis; gel electrophoresis; alkaline/Southern blotting; hybridization

Figure 1. A horizontal agarose gel apparatus. Wells are formed in the gel by placing a comb in the melted agarose before it hardens. The gel is then placed in a horizontal electrophoresis chamber filled with electrophoresis buffer and connected to a power supply. When voltage is applied to the system, the DNA migrates towards the anode in a tight ‘lane’ as shown in the diagram of the resulting gel.
Figure 2. The Southern transfer apparatus. As the buffer travels up the filter paper wick through the layers of filter paper, gel, membrane and paper towels, the DNA is deposited from the gel to the membrane. The weight (say, a textbook) ensures that all of the layers remain in close contact during the transfer process.
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 Further Reading
    book Ausubel FM, Brent R, Kingston RE et al. (eds) (1993) Current Protocols in Molecular Biology. New York: Wiley.
    book Old R and Primrose S (1994) Principles of Gene Manipulation, An Introduction to Genetic Engineering, 5th edn. London: Blackwell Scientific.
    book Sambrook J, Tritsch EF and Maniatis T (1989) Molecular Cloning. A Laboratory Manual. New York: Cold Spring Harbor Laboratory.
    Southern EM (2000) Blotting at 25. Trends in Biochemical Sciences 25(12): 585–588.
    book Westermeier R (2001) Electrophoresis in Practice, 3rd edn. New York: Wiley.

Related Sub-Topics:

Techniques and Tools in Clinical Genetics | Techniques and Tools in Molecular Biology | Biochemical and Biophysical Techniques
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Article Title: Electrophoresis and Blotting of DNA
 
How to Cite close
Tamber, Sandeep, and Hancock, Robert EW(Mar 2004) Electrophoresis and Blotting of DNA. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003746]