Dna Labeling

1. Dilute 500 ng sonicated genomic DNA to a final volume of 34.4 |l with distilled water; use amber 0.5-ml reaction tubes.

2. Add 32 |l of 2.5 x Random primer (mix before use), vortex 5-10 s and spin down.

3. Place the reaction for 10 min at 100°C (boiling water).

4. Place the reaction on ice for at least 5 min to ensure complete cooling of the reaction.

5. Spin down before adding the next components:

4.0 |l Cy-dye (Cy3 for patient DNA, Cy5 for reference DNA) 8.0 |l dNTPmix, vortexed before adding 1.6 |l Klenow fragment DNA polymerase

6. Vortex the reactions gently and spin down.

7. Incubate the reactions overnight in a 37°C water bath.

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