title: IGG_Mouse_178_AP1
status: Public on Jan 05 2007
contact.city: Genoa
contact.first: Paolo
contact.last: Fardin
contact.email: paolofardin@ospedale-gaslini.ge.it
contact.address: Largo G. Gaslini 5
contact.department: Molecular Biology
contact.country: Italy
contact.institute: Istiuto Giannina Gaslini
organism: Mus musculus
manufacture_protocol: C6-amino-linker oligonucleotides (50 nucleotides in length) were obtained from MWG oligoset (MWG-Biotech AG, Ebersberg, Germany), and spike-in controls oligonucleotides were purchased from Ambion ArrayControl Spot (Ambion Inc., Austin TX). Oligonucleotides were printed on e-Surf Activated Slides (Life Line Lab S.r.l., Italy) with a SpottingArray 24 (PerkinElmer, Wellesley, MA) using 4 Stealth Micro Spotting Pins (Telechem International, Inc. Sunnyvale, CA) in 150 mM phosphate buffer pH 8,5 at 40% humidity. E-surf activated slides are obtained by adsorption on glass of a hydrophilic polymer containing N,Nacryloyloxysuccinimide (NAS). Oligonucleotides were printed at a final concentration of 10 pmol/ml. The coupling reaction was performed o/n in a saturated NaCl solution chamber with a 75% relative humidity. All oligonucleotides were printed in quadruplicates over 4 subarrays with a 2x2 print head. Spike-in controls oligonucleotides, negative control and buffer were printed in quadruplicates onto each subarray. The scheme is repeated 3 times on the entire slide surface resulting in 12 replicates for each gene element and 48 replicates for each control element.