Comparative genomic hybridization using DNA from laser capture microdissected tissue.
Comparative genomic hybridization (CGH) is a powerful screening technique that can identify regions of gain and loss within the whole genome in a single experiment. The combination of laser capture microdissection, whole-genome amplification, and CGH permits genomic screening with high specificity and sensitivity. This complement of techniques has enabled analysis of focal regions and subpopulations of cells within a tissue, which has previously been difficult, providing insight into disease progression and heterogeneity. This chapter outlines the techniques involved in producing labeled probes from DNA extracted from laser capture microdissected material and the methods for hybridization of these probes to metaphase chromosomes. This protocol can also be applied to the preparation of probes for CGH arrays.
Callagy
Grace
G
Department of Oncology, University of Cambridge, UK.
Jackson
Lucy
L
Caldas
Carlos
C
D004273:DNA, Neoplasm / Q000032:analysis; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D009369:Neoplasms / Q000737:chemistry / Q000235:genetics / Q000473:pathology; D009693:Nucleic Acid Hybridization / Q000379:methods*
D004273:DNA, Neoplasm