Printing On Glass Slides

1. Array the PCR products from 384-well microtiter plates onto micro-slides (model Super Aldehyde substrates; Telechem International Inc., Sunnyvale, CA) using a microarray stamping machine (model SPBI02000; Hitachi Software Engineering Co., Ltd., Tokyo, Japan). The tip loads 2 |l of PCR products (500 to 1000 ng |l-1) from 384-well microtiter plates and deposits 0.5 nl per slide on 48 slides with spacing of 300 |m in our system (10).

2. Postprocess the slides according to the manufacturer's protocol (Telechem International Inc., Sunnyvale, CA). Dry the slides for more than 12 h in a desiccator (relative humidity <30%). This period may facilitate binding of the printed DNA and slide coating. Irradiate the slides with 65 mJ UV to obtain cross-linked DNA.

3. Rock them in 0.2% SDS for 2 min twice and then rock in distilled water for 2 min twice vigorously.

4. Transfer the slide racks into a chamber containing boiling water and leave for 2 min. Remove the slide racks to a clean glass container and leave them at room temperature for 5 min to cool. Pour the blocking solution, containing 1 g of sodium borohydride, 300 ml of phosphate-buffered saline (PBS; Invitrogen, Carlsbad, CA), and 90 ml of 100% ethanol, into the glass chamber.

5. Shake the slide racks gently for 5 min, transfer three times into a new chamber containing 0.2% SDS and shake gently for

6. Transfer them into a chamber containing distilled water, shake gently for 1 min, and dry by centrifugation for 20 min to remove any residual solution from the slides. Store the slides in a desiccator.

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