title: Chick Pineal 2004
status: Public on Jul 15 2004
contact.city: College Station
contact.first: Phillip
contact.middle: D.
contact.last: Beremand
contact.email: beremand@mail.bio.tamu.edu
contact.state: TX
contact.department: Biology
contact.country: USA
contact.institute: Texas A&M University
contact.phone: 979-845-0663
organism: Gallus gallus
description: The cDNA microarrays were prepared based upon approximately 4,500 PCR products from each of the two chicken pineal cDNA libraries constructed, for a total of ~9,000 ESTs. Briefly, the cDNA in plasmids from each library were PCR amplified using flanking primers (SK and T7), purified by ethanol precipitation using ammonium acetate, and placed in the wells of 96 well plates at a concentration of 50 ug/ml in 3X SSC. A GeneMachines OmniGrid microarrayer equipped with 8 Telechem SMP3 pins was used to spot the samples onto poly-L-lysine coated slides (CEL Associates). Duplicate 100 micron diameter spots were placed at intervals of 190 microns (center to center). Slides were printed in batches of 50–100. Printing was accomplished at 70 C and 60% humidity. The OmniGrid’s operating software used information on the position of the individual
  clones within the 96-well sample plates and the order in which the plates were used to create a file describing the position of each cDNA clone on the array for later analysis.
  Slides were stored desiccated at room temperature until their subsequent use. Prior to hybridization, the dried spots were hydrated gently over a steaming water bath, snap dried, and the DNA was UV cross-linked using a Stratalinker (Stratagene).
  Keywords = chick pineal gland
  Keywords = circadian oscillator