Protocol 33 Microarray hybridization and scanning

1. Spin the target sample for 1 min at 15 000 g to pellet any particulate matter. Pipette the target solution close to the end of the microarray slide.

2. Place a cover slip over the probe in such a way as to avoid the formation of bubbles.

3. Place the slides in a sealed hybridization cassette (Telechem International Inc., Sunnyvale, CA) and submerge in a 65°C water bath for 16-20 h.

4. After hybridization, wash the slides in 2 x SSC, 0.03% SDS for 2 min, then in 1 x SSC for 2 min, and finally in 0.05 x SSC for 2 min.

5. Centrifuge (1 min at 2500 g) the slides immediately to dry.

6. Scan the slides with a ScanArray 4000 (GSI Lumonics, Oxnard, CA) as described previously (9).

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