title: Gillichthys mirabilis cDNA microarray (Last Annotated 2/2010 by Cheryl A. Logan)
status: Public on Jul 15 2010
contact.city: Pacific Grove
contact.first: Cheryl
contact.middle: A
contact.last: Logan
contact.email: clogan@stanford.edu
contact.state: CA
contact.address: 120 Ocean View Blvd
contact.country: USA
contact.institute: Stanford 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 37C. cDNA inserts were amplified by PCR using 1 ul of bacterial suspension in 50 ul 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.