title: Assaying chromosomal inversions by single-molecule haplotyping.
abstract: Inversions are an important form of structural variation, but they are difficult to characterize, as their breakpoints often fall within inverted repeats. We have developed a method called 'haplotype fusion' in which an inversion breakpoint is genotyped by performing fusion PCR on single molecules of human genomic DNA. Fusing single-copy sequences bracketing an inversion breakpoint generates orientation-specific PCR products, exemplified by a genotyping assay for the int22 hemophilia A inversion on Xq28. Furthermore, we demonstrated that inversion events with breakpoints embedded within long (>100 kb) inverted repeats can be genotyped by haplotype-fusion PCR followed by bead-based single-molecule haplotyping on repeat-specific markers bracketing the inversion breakpoint. We illustrate this method by genotyping a Yp paracentric inversion sponsored by >300-kb-long inverted repeats. The generality of our methods to survey for, and genotype chromosomal inversions should help our understanding of the contribution of inversions to genomic variation, inherited diseases and cancer.
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SA, UK.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Turner
authors.forename: Jay
authors.initials: J
authors.lastname: Shendure
authors.forename: Greg
authors.initials: G
authors.lastname: Porreca
authors.forename: George
authors.initials: G
authors.lastname: Church
authors.forename: Peter
authors.initials: P
authors.lastname: Green
authors.forename: Chris
authors.initials: C
authors.lastname: Tyler-Smith
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Hurles
mesh_terms: D007446:Chromosome Inversion / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D020022:Genetic Predisposition to Disease / Q000235:genetics; D015894:Genome, Human / Q000235:genetics*; D006239:Haplotypes / Q000235:genetics*; D006467:Hemophilia A / Q000235:genetics; D006801:Humans; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*