title: Gillichthys mirabilis cDNA microarray
status: Public on Oct 01 2007
contact.city: Portland
contact.first: Brad
contact.last: Buckley
contact.state: OR
contact.address: 1719 SW 10th Ave
contact.department: Biology
contact.country: USA
contact.institute: Portland State University
organism: Gillichthys mirabilis
manufacture_protocol: This microarray consists of spotted cDNAs, generated by reverse transcribing mRNA collected from several tissues of the goby Gillichthys mirabilis. RNA was collected from liver, gill, brain and muscle tissue, extracted from individuals that had been exposed to heat, hypoxia or osmotic stress. cDNAs were directionally cloned into pTriplEx2 vector (Clontech; Mountain View, CA, USA). Plasmid libraries were transformed into E. coli and bacterial colonies were picked at random into 384-well microtiter plates and grown overnight at 37˚C. cDNA inserts were amplified by PCR using 1 μl of bacterial suspension in 50 μl standard PCR reactions with vector specific primers. After ethanol precipitation, and one round of washing in 70% ethanol, PCR products were prepared in 3X sodium citrate (SSC) and 1.5 mol-1 betaine, for spotting onto Fisher Scientific (Hampton, NH, USA) Gold Seal glass microscope slides. Slides were coated with poly-L-lysine.