title: Array-MAPH: a methodology for the detection of locus copy-number changes in complex genomes.
abstract: High-throughput genome-wide screening methods to detect subtle genomic imbalances are extremely important for diagnostic genetics and genomics. Here, we provide a detailed protocol for a microarray-based technique, applying the principle of multiplex amplifiable probe hybridization (MAPH). Methodology and software have been developed for designing unique PCR-amplifiable sequences (400-600 bp) covering any genomic region of interest. These sequences are amplified, cloned and spotted onto arrays (targets). A mixture of the same sequences (probes) is hybridized to genomic DNA immobilized on a membrane. Bound probes are recovered and quantitatively amplified by PCR, labeled and hybridized to the array. The procedure can be completed in 4-5 working days, excluding microarray preparation. Unlike array-comparative genomic hybridization (array-CGH), test DNA of specifically reduced complexity is hybridized to an array of identical small amplifiable target sequences, resulting in increased hybridization specificity and higher potential for increasing resolution. Array-MAPH can be used for detection of small-scale copy-number changes in complex genomes, leading to genotype-phenotype correlations and the discovery of new genes.
authors.affiliation: Department of Cytogenetics, The Cyprus Institute of Neurology & Genetics, Nicosia, Cyprus.
authors.forename: Ludmila
authors.initials: L
authors.lastname: Kousoulidou
authors.forename: Katrin
authors.initials: K
authors.lastname: Männik
authors.forename: Carolina
authors.initials: C
authors.lastname: Sismani
authors.forename: Olga
authors.initials: O
authors.lastname: Zilina
authors.forename: Sven
authors.initials: S
authors.lastname: Parkel
authors.forename: Helen
authors.initials: H
authors.lastname: Puusepp
authors.forename: Neeme
authors.initials: N
authors.lastname: Tõnisson
authors.forename: Priit
authors.initials: P
authors.lastname: Palta
authors.forename: Maido
authors.initials: M
authors.lastname: Remm
authors.forename: Ants
authors.initials: A
authors.lastname: Kurg
authors.forename: Philippos C
authors.initials: PC
authors.lastname: Patsalis
mesh_terms: D004247:DNA / Q000378:metabolism*; D023281:Genomics / Q000379:methods*; D046228:Microarray Analysis / Q000379:methods*; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012984:Software
chemical_list: D015335:Molecular Probes; D004247:DNA