[1]. Array Description: This microarray contains 4608 array elements spotted in duplicate. Since the microarray spotter is equipped with 12 capillary pins, the spots are arrayed over the entire length of the slide in 1 column of 12 blocks each containing 32x12 unique spots. The duplicates spots are printed in an identical column next to the first one. Spotting time to print 36 slides takes approximately 4 hours.
[2]. Array protocol description: Equipment: 1. Generation III microarray spotter (Amersham Biosciences, Buckinghamshire, UK). 2. 384 microtiter plates (Amersham Biosciences, Buckinghamshire, UK). 3. UV-crosslinker (Biolink, BLX-254, Vilber Lourmat, France). 4. Type VII silane coated slides (cat#: RPK0174, Amersham BioSciences, Buckinghamshire, UK). Spotting: 1. Dissolve cDNA array elements in 50% DMSO at concentrations ranging between 100-200ng/µl and array in 384 microtiter plates. 2. Set spotter settings for 4608 spots in duplicate: normal mode, 55% humidity, spot diameter: 250, spot buffer: 30, and pen deflection:0.508mm. 3. Make sure room temperature is between 20-24 degrees Celsius. 4. Place microtiter plates in the plate hotel with A1 in upper right corner. 5. Place 36 slides on the slide-deck and perform 2 manual pin-washes. 6. After spotting, let slides dry at 55% humidity for 3 hours and cross-link at 50 mjoules. 7. Block slides in %SSPE for 30 minutes at room temperature rinse in water and dry in centrifuge. 8. Slides can be stored for at least 6 months in a dry-cabinet.
