PNA Reaction

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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