1. The PNA probes are supplied ready to use in hybridization buffer (Applied Biosystems, Foster City, CA). Each PNA probe consists of a mixture of several short synthetic sequences (15-22 base units) specific for the centromeric repeated DNA sequence of the targeted chromosome. PNA probes specific for chromosomes 1 and 16 are used in the present protocol (see Note 3). The chromosome 1-specific probe is labeled in blue with diethlaminocoumarine. The chromosome 18-specific probe is labeled in red with rhodamine.
2. Phosphate-buffered saline (PBS; Gibco BRL, Eragny, France).
3. Tween-20 (Roche Diagnostics).
4. Washing buffers: 1X PBS, 0.1% Tween-20 and 2X SSC, 0.1% Tween-20.
5. 1.5 mL of sterile microcentrifuge tubes (Eppendorf AG).
6. Rubber cement (Artos, Strasbourg, France).
9. Humidified hybridization chamber.
11. Incubator set at 37°C.
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