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Fluorescence in situ hybridization to detect DNA sequences
FISH can also used to detect specific DNA sequences or gene loci (4, 15). To make DNA accessible to the probe, the cellular DNA must be denaturated by incubation in denaturation buffer.
Equipment and reagents
- Glass coverslips, 22x22mm
- 6-well plates
- 4% freshly made paraformaldehyde (Electron Microscopy Sciences) in PBS, pH 7.4
- Triton X-100 (Sigma)
- Parafilm
- Forceps, Dumont, GG (Electron Microscopy Sciences)
- Avidin-DCS-Texas Red or –fluorescein (Vector)
- Filterpaper
- Mounting medium (Molecular Probes)
- Microscopy coverslide
- Deionized formamide (Ambion)