title: Human and mouse oligonucleotide-based array CGH.
abstract: Array-based comparative genomic hybridization is a high resolution method for measuring chromosomal copy number changes. Here we present a validated protocol using in-house spotted oligonucleotide libraries for array comparative genomic hybridization (CGH). This oligo array CGH platform yields reproducible results and is capable of detecting single copy gains, multi-copy amplifications as well as homozygous and heterozygous deletions as small as 100 kb with high resolution. A human oligonucleotide library was printed on amine binding slides. Arrays were hybridized using a hybstation and analysed using BlueFuse feature extraction software, with >95% of spots passing quality control. The protocol allows as little as 300 ng of input DNA and a 90% reduction of Cot-1 DNA without compromising quality. High quality results have also been obtained with DNA from archival tissue. Finally, in addition to human oligo arrays, we have applied the protocol successfully to mouse oligo arrays. We believe that this oligo-based platform using 'off-the-shelf' oligo libraries provides an easy accessible alternative to BAC arrays for CGH, which is cost-effective, available at high resolution and easily implemented for any sequenced organism without compromising the quality of the results.
authors.affiliation: Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Paul
authors.initials: P
authors.lastname: van den Ijssel
authors.forename: Marianne
authors.initials: M
authors.lastname: Tijssen
authors.forename: Suet-Feung
authors.initials: SF
authors.lastname: Chin
authors.forename: Paul
authors.initials: P
authors.lastname: Eijk
authors.forename: Beatriz
authors.initials: B
authors.lastname: Carvalho
authors.forename: Erik
authors.initials: E
authors.lastname: Hopmans
authors.forename: Henne
authors.initials: H
authors.lastname: Holstege
authors.forename: Dhinoth Kumar
authors.initials: DK
authors.lastname: Bangarusamy
authors.forename: Jos
authors.initials: J
authors.lastname: Jonkers
authors.forename: Gerrit A
authors.initials: GA
authors.lastname: Meijer
authors.forename: Carlos
authors.initials: C
authors.lastname: Caldas
authors.forename: Bauke
authors.initials: B
authors.lastname: Ylstra
mesh_terms: D000818:Animals; D002460:Cell Line; D002869:Chromosome Aberrations; D002872:Chromosome Deletion; D004247:DNA / Q000302:isolation & purification; D023281:Genomics / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D004247:DNA