title: NMC_Rat_5k_cDNA_Ser2
status: Public on Mar 07 2003
contact.city: Trondheim
contact.first: Fekadu
contact.last: Yadetie
contact.email: fekadu.yadetie@mbi.uib.no
contact.address: Olav Kyrres gt 3
contact.department: Cancer Research and Molecular Medicine
contact.country: Norway
contact.institute: Norwegian University of Science and Technology
organism: Rattus norvegicus
description: The 5k rat cDNA arrays used here are part of a larger collection of 14k cDNAs purchased from Research Genetics (Huntsville, AL).   (http://nova.idi.ntnu.no/MCF/products.html). 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 prepared in 50% DMSO (about 50-200 ng/ l) and spotted onto 25 x 75mm amino-silane coated CMT-GAPS glass slides (Corning, NY) using a printing robot built in-house in collaboration with NEMKO (Trondheim, Norway) after a prototype developed at the National Human Genome Research Institute (NHGRI), (http://www.nhgri.gov/DIR/LCG/15K(HTML/). The arrays were spotted with 8 pins and contained 2 x 4 blocks of 31 x 31 spots. The probes were fixed to the slide surface by UV irradiation. The 5k cDNA probes represent 4908 unique UniGene clusters, of which 2511 cDNAs (51%) were 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 7688 elements including duplicates and controls.