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MORIMASA MATS UTA AND MAYUMI MATSUTA
etc., can be detected in interphase nuclei by dual-color FISH
using a combination of chromosome-specific and genespecific probes .
• CGH provides an overview of DNA sequence copy number
changes (losses, deletions, gains, amplifications) in a tumor
specimen and maps these changes on normal chromosomes,
in a single hybridization. CGH is in particular useful for the
analysis of DNA sequence copy number changes in common
solid tumors where complex karyotypes with numerous markers, double minutes, and homogeneously stained chromosomal regions are common. In other words, this technique
is useful for screening both unknown and known oncogenes.
Application of CGH in diagnosis and prognostic evaluation of
tumors is promising.
Others
The quantification and/or cell cycle analysis ofFISH -samples via
flow cytometry has been reported. Three-dimensional organization of chromosomes in interphase nuclei, which may change
with the cell cycle, can be analyzed using a confocal laser scanning microscope and image analysis devices.
Subprotocol 1
Fluorescence In Situ Hybridization (FISH)
These protocols have demonstrated the ability to detect hybridization of probes with target size of approx. 3 kb to whole genomes.
Outline
A short protocol of the entire FISH procedure is given here:
1. Denature specimen slides in 70% formamide12X SSC at 74°C
for 5 mins.