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Regeneration of comparative genomic hybridization oligonucleotide microarrays with dimethylurea.

Analytical biochemistry (2012-04-17)
Anna Ronowicz, Magdalena Brzeskwiniewicz, Piotr Madanecki, Patrick G Buckley, Ewa Orłowska, J Renata Ochocka, Janusz Limon, Arkadiusz Piotrowski
RESUMO

Reuse of materials in DNA hybridization-based methods has been known since the advent of Southern membranes. Array-based comparative genomic hybridization is essentially Southern hybridization with multiple probes immobilized on a solid surface. We show that comparative genomic hybridization microarrays fabricated with maskless array synthesizer technology can be used up to four times with the application of 1,3-dimethylurea as an array-stripping agent. We reproducibly detected chromosomal aberrations (0.6-22.4Mb in size) in four hybridization rounds using regenerated microarray slides. We also demonstrated that regenerated arrays can detect smaller alterations (16-200kbp), such as common copy number variants, as well as complex aberration profiles in tumor DNA.

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Sigma-Aldrich
N,N′-Dimethylurea, (sym.), ≥99% (from N)
Sigma-Aldrich
N,N′-Dimethylurea, (sym.), ≥95.0% (HPLC), technical
Supelco
N,N′-Dimethylurea, PESTANAL®, analytical standard