title: DGC v1.0
status: Public on Jul 16 2003
contact.city: Cambridge
contact.first: Colin
contact.last: Meiklejohn
contact.email: meiklej@fas.harvard.edu
contact.state: MA
contact.address: 16 Divinity Ave
contact.department: Organismic and Evolutionary Biology
contact.country: USA
contact.web_link: http://www.oeb.harvard.edu/hartl/lab/
contact.institute: Harvard University
contact.phone: 617-496-5540
organism: Drosophila melanogaster
description: 5928 cDNAs from the Drosophila Gene Collection version 1.0 were amplified by PCR with universal primers and PCR products were confirmed by gel electrophoresis. Added to these was a set of 177 separately amplified controls, each of which was replicated from one to 16 times on the array.  Clones for which the PCR product was absent, faint, the incorrect size, or showed multiple bands failed PCR quality control.  The PCR products were purified with Multiscreen filter plates (Millipore), dried down, resuspended in 30?L 3X SSC, and mechanically spotted onto polylysine-coated glass slides.  The arrays were allowed to air dry following printing and stored in a dessicator.  Prior to blocking, arrays were rehydrated by placing them in a humid chamber for 5-7 minutes and then snap dried by placing the slide (array up) on a 80 °C heating block for 3-5 seconds.  DNA was then crosslinked to the slide with 60 mJ of UV radiation.  Slides were blocked by immersion in a blocking solution (6 g succinic anhydride, 335 mL 1-methyl-2-polypyrolidinone, 15 mL 1 M sodium borate) and shaken at room temperature for 20 minutes.  They were then washed in boiling water for 2 minutes, rinsed in 95% ethanol, and spun dry.