Figure 5. Schematic representation of comparative genomic hybridization experimental set up.
Test DNA is labeled with biotin and normal diploid control DNA is labeled with digoxigenin. They are then combined (with an excess of Cot-I DNA), denatured, briefly allowed to reanneal and hybridized to normal diploid metaphase spreads. After detection of the biotin with avidin-FITC and the digoxigenin with mouse antidigoxigenin followed by goat antimouse TRITC, the metaphases are imaged, the ratio of green to red fluorescence intensity along the length of each chromosome plotted and the gain/loss profiles generated.
