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Mapping translocation breakpoints by next-generation sequencing

Item Type:Article
Title:Mapping translocation breakpoints by next-generation sequencing
Creators Name:Chen, W. and Kalscheuer, V. and Tzschach, A. and Menzel, C. and Ullmann, R. and Schulz, M.H. and Erdogan, F. and Li, N. and Kijas, Z. and Arkesteijn, G. and Pajares, I.L. and Goetz-Sothmann, M. and Heinrich, U. and Rost, I. and Dufke, A. and Grasshoff, U. and Glaeser, B. and Vingron, M. and Ropers, H.H.
Abstract:Balanced chromosome rearrangements (BCRs) can cause genetic diseases by disrupting or inactivating specific genes, and the characterization of breakpoints in disease-associated BCRs has been instrumental in the molecular elucidation of a wide variety of genetic disorders. However, mapping chromosome breakpoints using traditional methods, such as in situ hybridization with fluorescent dye-labeled bacterial artificial chromosome clones (BAC-FISH), is rather laborious and time-consuming. In addition, the resolution of BAC-FISH is often insufficient to unequivocally identify the disrupted gene. To overcome these limitations, we have performed shotgun sequencing of flow-sorted derivative chromosomes using "next-generation" (Illumina/Solexa) multiplex sequencing-by-synthesis technology. As shown here for three different disease-associated BCRs, the coverage attained by this platform is sufficient to bridge the breakpoints by PCR amplification, and this procedure allows the determination of their exact nucleotide positions within a few weeks. Its implementation will greatly facilitate large-scale breakpoint mapping and gene finding in patients with disease-associated balanced translocations.
Keywords:Base Sequence, Chromosome Breakage, Chromosome Mapping, Mental Retardation, Molecular Sequence Data, DNA Sequence Analysis, Genetic Translocation
Source:Genome Research
Publisher:Cold Spring Harbor Laboratory Press
Page Range:1143-1149
Date:July 2008
Official Publication:https://doi.org/10.1101/gr.076166.108
PubMed:View item in PubMed

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