title: Ips_pini-midgut.3
status: Public on Nov 13 2003
contact.city: Reno
contact.first: Christopher
contact.middle: Ian
contact.last: Keeling
contact.email: ckeeling@mac.com
contact.state: NV
contact.address: 1664 North Virginia Street
contact.department: Department of Biochemistry/Mail Stop 330
contact.country: USA
contact.web_link: http://bioinformatics.unr.edu/beetle/
contact.institute: University of Nevada, Reno
contact.phone: 775-784-1734
organism: Ips pini
description: The Ips_pini-midgut.3 array is a cDNA-based glass microarray produced for Dr. Claus Tittiger's laboratory by the Nevada Genomics Center at the University of Nevada, Reno. The array features 1920 spots representing 574 tentatively unique genes chosen from an Ips pini (the North American pine engraver beetle) EST project of anterior midgut tissue from juvenile hormone III treated (20 h) males (Eigenheer, A. L., et al., Gene 316:127-136, 2003 PMID:14563559).  Pheromone biosynthesis occurs de novo in this tissue.  cDNA was amplified using vector specific primers from selected clones.  PCR products were resuspended in Micro Spotting Solution Plus (Arrayit) and spotted onto SuperAmine microarray substrates (Arrayit) using a Virtek SDDC2 Microarrayer with a 1 x 8 array of SMP5B Stealth Micro Spotting pins (Arrayit) printing subarrays of 8 x 10 spots with 550 µm spacing.  Spotting was replicated in triplicate on each substrate with replicate spots 6 mm apart.  DNA was bound to substrate by first rehydrating the array, flash drying on a hot plate, crosslinking with 120 mJ of UV light (Stratagene UV Stratalinker 1800), and then baking for 60 min at 80°C.  Clones were chosen to represent the longest clone of a cluster-contig.
  Keywords = North American pine engraver beetle
  Keywords = anterior midgut
  Keywords = juvenile hormone
  Keywords = pheromone biosynthesis
  Keywords = Coleoptera
  Keywords = Scolytidae