Analysis of Replicating Mitochondrial DNA by In Organello Labeling and Two-Dimensional Agarose Gel Electrophoresis.

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dc.contributor.authorHolt, Ian J
dc.contributor.authorKazak, Lawrence
dc.contributor.authorReyes, Aurelio
dc.contributor.authorWood, Stuart R
dc.contributor.orcidReyes Tellez, Aurelio [0000-0003-2876-2202]
dc.date.accessioned2019-01-21T13:03:25Z
dc.date.available2019-01-21T13:03:25Z
dc.date.issued2016
dc.description.abstractOur understanding of the mechanisms of DNA replication in a broad range of organisms and viruses has benefited from the application of two-dimensional agarose gel electrophoresis (2D-AGE). The method resolves DNA molecules on the basis of size and shape and is technically straightforward. 2D-AGE sparked controversy in the field of mitochondria when it revealed replicating molecules with lengthy tracts of RNA, a phenomenon never before reported in nature. More recently, radioisotope labeling of the DNA in the mitochondria has been coupled with 2D-AGE. In its first application, this procedure helped to delineate the "bootlace mechanism of mitochondrial DNA replication," in which processed mitochondrial transcripts are hybridized to the lagging strand template at the replication fork as the leading DNA strand is synthesized. This chapter provides details of the method, how it has been applied to date and concludes with some potential future applications of the technique.
dc.description.sponsorshipWe thank Drs. Pohjoismäki and Jacobs for the EM image of the replicating mitochondrial DNA molecule, and the Medical Research Council for funding.
dc.format.mediumPrint
dc.identifier.doi10.17863/CAM.35580
dc.identifier.eissn1940-6029
dc.identifier.issn1064-3745
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/288262
dc.languageeng
dc.publisherSpringer New York
dc.publisher.urlhttp://dx.doi.org/10.1007/978-1-4939-3040-1_8
dc.subject2D-AGE
dc.subjectDNA labeling
dc.subjectDNA replication
dc.subjectHumans
dc.subjectMammals
dc.subjectMitochondrial DNA
dc.subjectReplication intermediates
dc.subjectTwo-dimensional agarose gel electrophoresis
dc.subjectAnimals
dc.subjectCell Line, Tumor
dc.subjectDNA Replication
dc.subjectDNA, Mitochondrial
dc.subjectElectrophoresis, Gel, Two-Dimensional
dc.subjectHeLa Cells
dc.subjectHumans
dc.subjectLiver
dc.subjectMitochondria
dc.subjectMutation
dc.subjectNeurodegenerative Diseases
dc.subjectRats
dc.subjectStaining and Labeling
dc.titleAnalysis of Replicating Mitochondrial DNA by In Organello Labeling and Two-Dimensional Agarose Gel Electrophoresis.
dc.typeArticle
prism.endingPage113
prism.publicationDate2016
prism.publicationNameMethods Mol Biol
prism.startingPage95
prism.volume1351
rioxxterms.licenseref.startdate2016-01
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.typeJournal Article/Review
rioxxterms.versionNA
rioxxterms.versionofrecord10.1007/978-1-4939-3040-1_8
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