title: Chaetoceros neogracile 1.8 Kb cDNA microarray platform
status: Public on Dec 13 2007
contact.city: seoul
contact.first: Gyeongseo
contact.last: Jung
contact.address: 17 haengdang-dong, seongdong-gu
contact.country: South Korea
contact.institute: Hanyang university
organism: Chaetoceros neogracilis
manufacture_protocol: We constructed the cDNA microarrays from the Chaetoceros neogracile cDNA library (Jung et al., 2007). 2500 selected expressed sequence tags (ESTs) from each clone, including housekeeping genes like ribosomal binding proteins, were amplified by PCR using M13 primers and recovered by a MultiScreen PCR Filter Plate (Millipore, Billerica, MA, USA). Each cDNA chip was designed with 1774 successfully amplified PCR products from the cDNA library. Additionally, several control clones from the Saccharomyces cerevisiae intergeneic sequences were included. Microarrays were produced using a fully automated system by robotic operation at Digital Genomics (Seoul, South Korea). The cDNA PCR products were spotted onto amine-coated GAPSII slides (Corning Inc., Corning, NY, USA) using MG II (Biorobotics Inc., Woburn, MA, USA) with a 240 um pitch between spots as a twin format. There were 22 x 22 spots per block, and the spot diameter was 100~140 um. To evaluate fabrication, a cDNA microarray was stained with Syto61 (Molecular Probes, Eugene, OR, USA).