Genomic Unit, Biotechnology Program, Spanish National Cancer Research Centre
Manufacture protocol
1.- Individual I.M.A.G.E. Consortium [LLNL] clones were grown in 96-well plates at 37°C for 6 hours. 2.- The bacterial cultures were used for PCR amplification. 3.- PCR products were purified by ultrafiltration, verified by agarose gel electrophoresis and quantified by gel image densitometry. 4.- Probe concentrations were adjusted in 50% DMSO at about 100-150 ng/ul. 5.- Probes were printed onto Gamma Amino Silane Coated glass slides (CMT-GAPS Corning) using the spotter Multigrid II (BioRobotics). 6.- After printing, the slides were baked for 4 hours at 80ºC and stored desiccated in the dark, under vacuum and at room temperature.
Support
glass
Coating
γ-amino silane
Description
CNIO Human Oncochip 2.0 is a cDNA microarray which consists of 27648 features, with 11768 cDNA clones representing over 9300 genes. This microarray is specially designed to study the genes involved in cancer, it includes: 1) genes with known molecular function and involved in cell processes related to cancer like cell cycle control, apoptosis, angiongenesis, cell adhesion, etc.; 2) tissue specific genes; 3) housekeeping genes as positive controls and non human genes as negative controls. Since the microarray spotter is equipped with 48 capillary pins, the spots are arrayed over the entire length of the slide in 48 subarrays, arranged in 4 meta-columns and 12 meta-rows. Each subarrays consists 24 columns x 24 rows.
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