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Two-dimensional strandness-dependent electrophoresis.

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Authors
Gunnarsson, Gudmundur H
Gudmundsson, Bjarki
Thormar, Hans G
Alfredsson, Arni
Jonsson, Jon J
Issue Date
2007-01-01

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Citation
Nat Protoc 2007, 1(6):3011-8
Abstract
Two-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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http://dx.doi.org/10.1038/nprot.2006.477
ae974a485f413a2113503eed53cd6c53
10.1038/nprot.2006.477
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