Resuspending 1-1. Oligos from 20K set are arrayed by Operon in 384-well V-bottom plates (Genetix). Each well contains 1 nmol of oligo. 1-2. Resuspend oligos in water (20µM) on Staccato RapidPlate (Caliper). 1-3. Incubate at room temperature during a night shaking plates to aid resuspension of DNA. 1-4. Deep freeze and defrost centrifuging plates at 1000 rpm for 16 minutes then 2000 rpm for 3 minutes. 1-5. Take 5 µl of oligos solution and put it in new polypropylene v-bottom plates (cat# X7022, Genetix) 1-6. Add 5 µL of 1X Pronto! Universal Spotting Solution (Corning) to each well and mix. Spotting 2-1. Centrifuge plates at 2000 rpm for 3 minutes. 2-2. Oligos are ready for spotting. 2-3. Spotting on glass slides (Corning, Ultragaps), with 48 Microspotting pins Stealth 3 on Chipwriter (Virtek), with control of humidity (45-50%). 2-4. Fixation of the oligos slides with UV (600 mJoules). 2-5. Storage the slides in a vacuum. 2-6. After the print run is completed, cover oligos plates with seals and deepfreeze them at -20°C in a protected environment.
Support
glass
Coating
aminosilane
Description
Roslin/ARK Cooperative Research Chicken Array set. The oligo set (20,673) were designed by ArK using ENSEMBL genes transcripts, the BBSRC fully sequenced set and the DT40 full length sequencing set. This oligo set was synthesized by Operon (2007).
Gene expression profiling to identify uterine chicken proteins associated with the supply of minerals and potentially actives on the biomineralisation process during eggshell formation
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