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          Institute: MPI für molekulare Genetik     Collection: Department of Human Molecular Genetics     Display Documents



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ID: 469015.0, MPI für molekulare Genetik / Department of Human Molecular Genetics
Breakpoint analysis of balanced chromosome rearrangements by next-generation paired-end sequencing
Authors:Chen, Wei; Ullmann, Reinhard; Langnick, Claudia; Menzel, Corinna; Wotschofsky, Zofia; Hu, Hao; Döring, Andreas; Hu, Yuhui; Kang, Hui; Tzschach, Andreas; Hoeltzenbein, Maria; Neitzel, Heidemarie; Markus, Susanne; Wiedersberg, Eberhard; Kistner, Gerd; van Ravenswaaij-Arts, Conny M. A.; Kleefstra, Tjitske; Kalscheuer, Vera M.; Ropers, Hans-Hilger
Language:English
Date of Publication (YYYY-MM-DD):2010
Title of Journal:European Journal of Human Genetics
Journal Abbrev.:Eur J Hum Genet
Copyright:© 2010 European Society of Human Genetics
Review Status:not specified
Audience:Experts Only
Abstract / Description:Characterisation of breakpoints in disease-associated balanced chromosome rearrangements (DBCRs), which disrupt or inactivate specific genes, has facilitated the molecular elucidation of a wide variety of genetic disorders. However, conventional methods for mapping chromosome breakpoints, such as in situ hybridisation with fluorescent dye-labelled bacterial artificial chromosome clones (BAC-FISH), are laborious, time consuming and often with insufficient resolution to unequivocally identify the disrupted gene. By combining DNA array hybridisation with chromosome sorting, the efficiency of breakpoint mapping has dramatically improved. However, this can only be applied when the physical properties of the derivative chromosomes allow them to be flow sorted. To characterise the breakpoints in all types of balanced chromosome rearrangements more efficiently and more accurately, we performed massively parallel sequencing using Illumina 1G analyser and ABI SOLiD systems to generate short sequencing reads from both ends of DNA fragments. We applied this method to four different DBCRs, including two reciprocal translocations and two inversions. By identifying read pairs spanning the breakpoints, we were able to map the breakpoints to a region of a few hundred base pairs that could be confirmed by subsequent PCR amplification and Sanger sequencing of the junction fragments. Our results show the feasibility of paired-end sequencing of systematic breakpoint mapping and gene finding in patients with disease-associated chromosome rearrangements.
Free Keywords:Breakpoint mapping, Next-generation sequencing technology, Paired-end sequencing
Comment of the Author/Creator:email: wei.chen@mdc-berlin.de
email: kalscheu@molgen.mpg.de
External Publication Status:published
Document Type:Article
Communicated by:Hans-Hilger Ropers
Affiliations:MPI für molekulare Genetik
External Affiliations:Max-Delbrück-Centrum für Molekulare Medizin, Berlin Institute for Medical Systems Biology, Berlin, Germany
Institute of Human Genetics, Charité – Universitätsmedizin Berlin, Berlin, Germany
Praxis für medizinische Genetik Regensburg, Regensburg, Germany
HELIOS-Kliniken Schwerin, Schwerin, Germany
Labor für humangenetische Diagnostik, Schwerin, Germany
Department of Genetics, University Medical Center Groningen, University of Groningen, The Netherlands
Department of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
Identifiers:DOI:10.1038/ejhg.2009.211
ISSN:1018-4813
URL:http://www.nature.com/ejhg/journal/vaop/ncurrent/p...
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