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Technical Advances |
,
,¶,
From the Departments of Oncology *
and
Pathology,
the Cambridge Institute for
Medical Research/Wellcome Trust Centre for Molecular Mechanisms in
Disease,
and Strangeways Research
Laboratories,¶
University of Cambridge, Cambridge;
and the Cambridge Breast Unit,
Addenbrooke’s
Hospital, Cambridge, United Kingdom
We have developed a protocol for degenerate oligonucleotide-primed-polymerase chain reaction-based array comparative genomic hybridization (array CGH) that, when combined with a laser microdissection technique, allows the analysis of cancer cell populations isolated from routine, formalin-fixed, paraffin-embedded tissue samples. Comparison of copy number changes detected by degenerate oligonucleotide-primed-polymerase chain reaction-based array CGH to those detected by conventional array CGH or fluorescence in situ hybridization, demonstrated that amplifications can be reliably detected. Using a genomic microarray containing 57 oncogenes, we screened a total of 28 breast cancer samples and obtained a detailed amplicon profile that is the most comprehensive to date in human breast cancer. The array CGH method described here will allow the genetic analysis of paraffin-embedded human cancer materials for example in the context of clinical trials.
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