Agarose and Polyacrylamide Gel Electrophoresis
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Electrophoresis through agarose or polyacrylamide gels is a standard method used to separate, identify, and purify nucleic
acids. The technique is simple, rapid to perform and capable of resolving fragments that differ by as little as 0.2% in size.
Electrophoresis occurs under the influence of an electric field: Charged molecules such as nucleic acids migrate in the direction
of the electrode having the opposite charge (anode). The electrophoretic mobility of nucleic acids is determined by a number
of parameters, but molecules of linear double-stranded DNA migrate through gel matrices at rates that are inversely proportional
to the log10 of the number of base pairs (1) and therefore larger molecules migrate more slowly because of the greater frictional drag (see
Note 1). Other factors affecting electrophoretic mobility include the pK value, base composition, concentration of gel matrix, composition and ionic strength of the electrophoresis buffer, temperature
and the use of intercalating dyes such as ethidium bromide.
Affiliation(s): (3) Department of Haematology, Birmingham Children’s Hospital NHS Trust, Birmingham, UK
Book Title: PCR Mutation Detection Protocols
Series: Methods in Molecular Biology | Volume: 187 | Pub. Date: Apr-04-2002 | Page Range: 1-12 | DOI: 10.1385/1-59259-273-2:001
Subject: Genetics/Genomics
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