Human and mouse oligonucleotide-based array CGH.
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.
van den Ijssel
Paul
P
Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands.
Tijssen
Marianne
M
Chin
Suet-Feung
SF
Eijk
Paul
P
Carvalho
Beatriz
B
Hopmans
Erik
E
Holstege
Henne
H
Bangarusamy
Dhinoth Kumar
DK
Jonkers
Jos
J
Meijer
Gerrit A
GA
Caldas
Carlos
C
Ylstra
Bauke
B
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
D004247:DNA