title: Mapping translocation breakpoints by next-generation sequencing.
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.
authors.affiliation: Max Planck Institute for Molecular Genetics, 14195 Berlin, Germany. wei@molgen.mpg.de
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Vera
authors.initials: V
authors.lastname: Kalscheuer
authors.forename: Andreas
authors.initials: A
authors.lastname: Tzschach
authors.forename: Corinna
authors.initials: C
authors.lastname: Menzel
authors.forename: Reinhard
authors.initials: R
authors.lastname: Ullmann
authors.forename: Marcel Holger
authors.initials: MH
authors.lastname: Schulz
authors.forename: Fikret
authors.initials: F
authors.lastname: Erdogan
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.forename: Zofia
authors.initials: Z
authors.lastname: Kijas
authors.forename: Ger
authors.initials: G
authors.lastname: Arkesteijn
authors.forename: Isidora Lopez
authors.initials: IL
authors.lastname: Pajares
authors.forename: Margret
authors.initials: M
authors.lastname: Goetz-Sothmann
authors.forename: Uwe
authors.initials: U
authors.lastname: Heinrich
authors.forename: Imma
authors.initials: I
authors.lastname: Rost
authors.forename: Andreas
authors.initials: A
authors.lastname: Dufke
authors.forename: Ute
authors.initials: U
authors.lastname: Grasshoff
authors.forename: Birgitta
authors.initials: B
authors.lastname: Glaeser
authors.forename: Martin
authors.initials: M
authors.lastname: Vingron
authors.forename: H Hilger
authors.initials: HH
authors.lastname: Ropers
mesh_terms: D000293:Adolescent; D001483:Base Sequence; D002648:Child; D019457:Chromosome Breakage; D002874:Chromosome Mapping / Q000379:methods; D005260:Female; D006801:Humans; D008607:Intellectual Disability / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*; D014178:Translocation, Genetic