Chromosomal breakpoint mapping by arrayCGH using flow-sorted chromosomes.
Despite the recent completion of the human genome project, the mapping of disease-related chromosomal translocation breakpoints and genes has remained laborious. Here, we describe a novel and rapid procedure to map such translocation breakpoints using flow-sorted chromosomes in combination with array-based comparative genomic hybridization (arrayCGH). To test the feasibility of this approach, we used a t(12;15)(q13;q25)-positive cell line with known breakpoint positions as a model. The derivative 12 chromosomes were flow-sorted, labeled, and hybridized to a genome-wide array containing 3648 well-characterized human genomic clones. The exact locations of the breakpoints on both chromosome 12 and 15 could be determined in a single hybridization experiment. In addition, we have tested the minimal amount of material necessary to perform these experiments and show that it is possible to obtain highly reliable profiles using as little as 10,000 flow-sorted chromosomes.
Veltman
Imke M
IM
Department of Human Genetics, 417, University Medical Center Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands. i.veltman@antrg.umcn.nl
Veltman
Joris A
JA
Arkesteijn
Ger
G
Janssen
Irene M
IM
Vissers
Lisenka E
LE
de Jong
Pieter J
PJ
van Kessel
Ad Geurts
AG
Schoenmakers
Eric F
EF
D019457:Chromosome Breakage / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods*; D005240:Feasibility Studies; D005434:Flow Cytometry / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D007621:Karyotyping / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods