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dc.contributor.authorGunnarsson, Gudmundur H*
dc.contributor.authorGudmundsson, Bjarki*
dc.contributor.authorThormar, Hans G*
dc.contributor.authorAlfredsson, Arni*
dc.contributor.authorJonsson, Jon J*
dc.date.accessioned2008-03-17T14:45:58Z
dc.date.available2008-03-17T14:45:58Z
dc.date.issued2007-01-01
dc.date.submitted2008-03-17
dc.identifier.citationNat Protoc 2007, 1(6):3011-8en
dc.identifier.issn1750-2799
dc.identifier.pmid17406562
dc.identifier.doi10.1038/nprot.2006.477
dc.identifier.urihttp://hdl.handle.net/2336/20858
dc.descriptionTo access publisher version of this article. Please click on the hyperlink in Additional Links fielden
dc.description.abstractTwo-dimensional strandness-dependent electrophoresis (2D-SDE) separates nucleic acids in complex samples according to strandness, conformation and length. Under the non-denaturing conditions of the first electrophoretic step, single-stranded DNA, double-stranded DNA and RNA.DNA hybrids of similar length migrate at different rates. The second electrophoretic step is performed under denaturing conditions (7 mol l(-1) urea, 55 degrees C) so that all the molecules are single-stranded and separate according to length only. 2D-SDE is useful for revealing important characteristics of complex nucleic acid samples in manipulations such as amplification, renaturation, cDNA synthesis and microarray hybridization. It can also be used to identify mispaired, nicked or damaged fragments in double-stranded DNA. The protocol takes approximately 2 h and requires only basic skills, equipment and reagents.
dc.language.isoenen
dc.publisherNature Pub. Groupen
dc.relation.urlhttp://dx.doi.org/10.1038/nprot.2006.477en
dc.subject.meshBase Pair Mismatchen
dc.subject.meshDNA Breaks, Single-Strandeden
dc.subject.meshDNA Damageen
dc.subject.meshDNA, Single-Strandeden
dc.subject.meshElectrophoresis, Gel, Two-Dimensionalen
dc.subject.meshNucleic Acid Heteroduplexesen
dc.subject.meshResearch Designen
dc.titleTwo-dimensional strandness-dependent electrophoresis.en
dc.typeArticleen
dc.contributor.departmentDepartment of Biochemistry and Molecular Biology, Faculty of Medicine, University of Iceland, IS-101 Reykjavik, Iceland.en
dc.identifier.journalNature protocolsen
html.description.abstractTwo-dimensional strandness-dependent electrophoresis (2D-SDE) separates nucleic acids in complex samples according to strandness, conformation and length. Under the non-denaturing conditions of the first electrophoretic step, single-stranded DNA, double-stranded DNA and RNA.DNA hybrids of similar length migrate at different rates. The second electrophoretic step is performed under denaturing conditions (7 mol l(-1) urea, 55 degrees C) so that all the molecules are single-stranded and separate according to length only. 2D-SDE is useful for revealing important characteristics of complex nucleic acid samples in manipulations such as amplification, renaturation, cDNA synthesis and microarray hybridization. It can also be used to identify mispaired, nicked or damaged fragments in double-stranded DNA. The protocol takes approximately 2 h and requires only basic skills, equipment and reagents.


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