title: Host and Pathogen Microarray
status: Public on Sep 27 2007
contact.city: London
contact.first: Paul
contact.last: Kellam
contact.email: p.kellam@ucl.ac.uk
contact.state: London
contact.address: 46 Cleveland Street
contact.department: Infection
contact.country: United Kingdom
contact.web_link: http://www.ucl.ac.uk/medicalschool/infection-immunity/research/group-leaders/pkellam.htm
contact.institute: UCL
contact.phone: 00442076799559
organism: Homo sapiens
  Human gammaherpesvirus 8
manufacture_protocol: The purified DNA probes in spotting buffer were deposited onto aminopropyl silane-coated Nexterion Slide A (Schott, UK) using the Qarray2 arrayer (Genetix, UK) with a 12-pin print head of Genetix aQu 75um split pins (Genetix, UK).  Spotted arrays were air-dried for 12+ hours, followed by heating at 80 °C in dry oven for 3 h to immobilise the DNA.  Unbound surfaces on the arrays were inactivated by immersing in 60 ml blocking solution: 5× SSC, 0.5% (w/v) bovine serum albumin (BSA, Sigma, UK), 0.5% (w/v) non-fat powdered milk (Tesco, UK) at 50 °C for 45 min, followed by washing 2 times in deionised water.  Then the arrays were incubated in deionised water at 95 °C for 2 min to denature the DNA.  Denatured arrays were washed and blocked again (with fresh solution) at 50 °C for 30 min.  Finally the arrays were washed 2 times in deionised water, immersed in isopropanol (Sigma, UK) for 30 seconds and immediately dried by spinning at 200 g, 2 min.  Processed arrays were stored in a dry environment for use within 24 hours.