Abstract
Pulsed-field gel electrophoresis (PFGE) is a technique for resolving large (up to 10 Mb) DNA molecules. Using multiple pairs of electrodes DNA is subject to an alternating electric field through a solid agarose matrix. As the current changes direction the reorientation time of DNA is proportional to molecular weight; thus fragments are separated in the gel based on their size. Here we describe the use of PFGE to analyze DNA double-strand break formation and repair in human chromosomal DNA.
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References
Schwartz DC, Cantor CR (1984) Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell 37:67–75
Gardiner K, Laas W, Patterson D (1986) Fractionation of large mammalian DNA restriction fragments using vertical pulsed-field gradient gel electrophoresis. Somat Cell Mol Genet 12:185–195
Chu G, Vollrath D, Davis RW (1986) Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science 234:1582–1585
Saleh-Gohari N, Bryant HE, Schultz N, Parker KM, Cassel TN, Helleday T (2005) Spontaneous homologous recombination is induced by collapsed replication forks that are caused by endogenous DNA single-strand breaks. Mol Cell Biol 25:7158–7169
Bryant HE, Ying S, Helleday T (2006) Homologous recombination is involved in repair of chromium-induced DNA damage in mammalian cells. Mutat Res 599:116–123
Ying S, Myers K, Bottomley S, Helleday T, Bryant HE (2009) BRCA2-dependent homologous recombination is required for repair of Arsenite-induced replication lesions in mammalian cells. Nucleic Acids Res 37:5105–5113
Herschleb J, Ananiev G, Schwartz DC (2007) Pulsed-field gel electrophoresis. Nat Protoc 2:677–684
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Bryant, H.E. (2012). DNA Double-Strand Break Damage and Repair Assessed by Pulsed-Field Gel Electrophoresis. In: Bjergbæk, L. (eds) DNA Repair Protocols. Methods in Molecular Biology, vol 920. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-998-3_22
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DOI: https://doi.org/10.1007/978-1-61779-998-3_22
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-997-6
Online ISBN: 978-1-61779-998-3
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