title: NMC_Rat_15k_cDNA_Ser6
status: Public on Oct 20 2004
contact.city: Trondheim
contact.first: Kristin
contact.last: Nørsett
contact.email: kristin.norsett@medisin.ntnu.no
contact.department: Dept of Cancer Research and Molecular Medicine
contact.country: Norway
contact.institute: Norwegian University of Science and Technology
contact.phone: +47 73 59 88 02
organism: Rattus norvegicus
description: The 15k rat cDNA arrays were prepared from a collection of 14k cDNAs purchased from Research Genetics (Huntsville, AL) (http://www.mikromatrise.no). Plasmids were isolated from overnight bacterial cultures in 96-well plates and cDNA inserts amplified by PCR using M13 primers 5'-ctg caa ggc gat taa gtt ggg taa c and 5'-gtg agc gga taa caa ttt cac aca gga aag a. The PCR products were purified with Montage PCR purification kit (Millipore) and quality and concentration was assessed by agarose gel electrophoresis. The purified PCR products were dissolved in 50% DMSO (about 50-200 ng/ l) and spotted in duplicate onto 25 x 75mm amino-silane coated CMT-GAPS glass slides (Corning, NY) using a MicroGrid II printing robot (BioRobotics; Cambridge, UK). The arrays were spotted with 48 pins and contained 4 x 12 blocks of 25 x 25 spots. The probes were fixed to the slide surface by UV irradiation (200 mJ). The 15k cDNA arrays represent 14018 unique probes, all printed in duplicates. In addition, a few in-house cDNA probes and a set of positive and negative control elements including 10 different PCR products of exogenous DNA from SpotReport™ cDNA Array Validation System (Stratagene, La Jolla, CA) were printed on the arrays. Totally, each array contained 30000 elements including duplicates and controls.