title: SKLMVGE_TR_FC_V1.0
status: Public on Sep 23 2008
contact.city: Beijing
contact.first: Qi
contact.last: Jin
contact.email: zdsys@vip.sina.com.cn
contact.state: Beijing
contact.address: BDA Rong Jing east street No.6, 3rd floor
contact.department: National Institute for Viral Disease Control and Prevention
contact.country: China
contact.institute: Chinese Center for Disease Control and Prevention
contact.phone: +86(010)67876915
organism: Trichophyton rubrum
description: In this chip there were two fields named A and B, they described the upper and lower part (location) of the chip. Each field contains 10,000 spots. The spots in field A are same with those in field B.
  
manufacture_protocol: Materials are transferred from microtiter plate wells to the slide surface by an automated print head
  The cDNA libraries were constructed following the protocols of the SUPERSCRIPTTM Plasmid System with GATEWAYTM Technology for cDNA Synthesis and Cloning (Invitrogen).  The cDNA plasmids were isolated by the Millipore method using MADV filter plates.  Sequencing was performed with a generic T7 primer located 5' upstream of the inserted segments, following the protocol for the PRISM Big Dye terminator kit on an ABI 3700 automated sequencer(Applied Biosystems, Foster City, CA).
  EST processing pipeline and annotation
  Phred quality assessment and computational analysis were carried out as described previously [4].
  cDNA microarrays
  PCR fragments used for printing the microarray chip were amplified from the ESTs library in 96-well plates using vector-PCR amplification with T7 and SP6 universal primers.  PCR products were analyzed on gels to confirm the success of the reactions and were subsequently purified using MultiScreen-PCR plates (Millipore Corporation, Bedford, USA).  Purified PCR products were resuspened in 12 ¦Ìl of spotting solution containing 50% DMSO to produce the microarrays.  A set of microarrays containing a total of 10,000 spots (including 8,997 clones in the form of PCR products, and 1,003 controls in each block, including blank, negative, and positive controls) were spotted in duplicated onto coated glass slides (Corning, NY) with a Cartesian arrayer.