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Mutagenesis 2006 21(4):243-247; doi:10.1093/mutage/gel022
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© The Author 2006. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Detection of Alu sequences and mtDNA in comets using padlock probes

Sergey Shaposhnikov1, Chatarina Larsson2, Sara Henriksson2, Andrew Collins1,* and Mats Nilsson2

1 Institute of Basic Medical Sciences, University of Oslo PB 1046 Blindern, 0316 Oslo, Norway 2 Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University Se-75185 Uppsala, Sweden

Single cell gel electrophoresis, or the comet assay, is widely used to measure DNA damage and repair. However, the behaviour of the DNA under the conditions used for the comet assay is not fully understood. In developing a method for studying specific gene sequences within comets, using ‘padlock probes’ (circularizable oligonucleotide probes), we have first applied probes that hybridize to Alu repetitive elements and to mitochondrial DNA (mtDNA). During the sequence of stages in the comet assay, mtDNA progressively disperses into the surrounding agarose gel, showing no tendency to remain with nuclear DNA in the comets. In contrast, Alu probes remain associated with both tail and head DNA.

*To whom correspondence should be addressed: Department of Nutrition, University of Oslo, POB 1046 Blindern, 0316 Oslo, Norway; Tel: +47 22 85 13 60; Fax: +47 22 85 13 41; E-mail: andrew.collins{at}medisin.uio.no


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