title: KCL-Danio_rerio-16K-pronto-v1.0
status: Public on May 20 2010
contact.city: Cardiff
contact.first: Peter
contact.last: Kille
contact.email: kille@cardiff.ac.uk
contact.address: Park Place
contact.department: Cardiff School of Biosciences
contact.country: United Kingdom
contact.web_link: http://kille.cf.ac.uk
contact.institute: Cardiff University
contact.phone: 02920874507
organism: Danio rerio
manufacture_protocol: Oligonucleotides include 16,399 designed and synthesised by Compugen and Sigma Genesys as a Zebrafish OligoLibrary ready set, which represent 15,806 LEADS™ clusters plus 171 controls (http://www.labonweb.com/chips/libraries.html). In addition, 331 customised oligonucleotides and 23 controls from Amersham Lucidea Universal ScorecardTM were added to the array set.
  All oligonucleotides were resuspended to a concentration of 25uM in Pronto buffer (Corning).
  The Zebrafish oligonucleotides arrays were spotted on UltraGAPS™ coated slides (Corning Life Sciences, Promega), using 48 SMP3 Telechem split pins mounted  Qarray2 robot (Genetix Ltd) .
  Printing conditions include: Humidity range - 40 – 50 %, temperature range = 20 – 25 ºC, Inking time: 3 sec, Inking depth: about 1mm below the solution surface, Printing depth: adjust to high enough to make all pins just touch, Stamping time (dwell time): 50msec, Maximum number of prints per inking: up to 400 without losing intensity but avoid re-inking, Spot diameter = 100 microns, Spot spacing = 215 microns, five blots are performed prior to printing, pins are washed using 3x5sec water wash and 2x5sec ethanol wash between sample printing.
  Immobilization of the cDNA onto the Ultra-GAPS coated glass slides was achieved by means of baking and UV crosslinking. The printed slides were initially immobilized by placing them in a UV StratalinkerTM 2400 bench top transilluminator (Stratagene Ltd)
  The slides were then placed in a lightproof container and baked in an oven at 80oC for 2 hours. The slides were then stored in a lightproof desiccator at room temperature, until required.