Hybridization

1. Boil the probe for 3 min by suspending the tube in boiling water and quench on ice. Check the volume after boiling and restore to the initial volume with sterile distilled water.

2. Pipet 20 | L of the probe onto the chromosomes. Cover the chromosomes and probe with a clean siliconized cover slip. There is no need to seal the cover slip.

3. Place the slides in a plastic box lined with moist tissue to prevent evaporation from the preparation, seal the lid, and place in a 58°C incubator overnight.

4. Remove the slides from the humid box. Dip them in 2X SSC to allow the cover slip to slide off, then wash them in 2X SSC, 53°C, for 1 h. Three changes of wash solution should be made.

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