title: UGI Human 14112 V2.0
status: Public on Sep 25 2008
contact.city: Shanghai
contact.first: Mei
contact.last: Jiang
contact.email: 042023070@fudan.edu.cn
contact.address: Handan Road
contact.department: Institute of Genetics
contact.country: China
contact.institute: Fudan University
organism: Homo sapiens
manufacture_protocol: The construction of the microarrays used in this study was carried out following Brown’s method (available at http://cmgm.stanford.edu/pbrown/protocols/index.html).We fabricated a “genome-wide” cDNA microarray consisting of 14061 sequences including full-length and partial cDNAs representing novel, known, and control genes provided by United Gene Holdings Group Co., Ltd. Genes represented in the arrays were identified with high similarity to the sequences in Unigene database of NCBI by performing Blast (http://www.ncbi.nlm.nih.gov/).The cDNA inserts were amplified by polymerase chain reaction (PCR) with universal primers to plasmid vector sequences and then purified. All the PCR products were examined by agarose gel electrophoresis to ensure the quality and identity of the amplified clones as expected. Then the amplified PCR products were dissolved in a buffer containing 3ХSSC solution. The solution with amplified PCR products were spotted onto silylated slides(CEL Associates, Houston, TX) using a Cartesian PixSys 7500 motion control robot (Cartesian Technologies, Irvine, CA) fitted with ChipMaker Micro-Spotting Technology (TeleChem International, Sunnyvale, CA). Glass slides with spotted cDNA were then hydrated for 2 hr in 70% humidity, dried for 0.5 hr at room temperature, and UV cross-linked (65mj/cm). They were further processed at room temperature by soaking in 0.2% sodium dodecyl sulfate (SDS) for 10 min, distilled H2O for 10 min, and 0.2% sodium borohydride (NaBH4) for 10 min. The slides were dried again and ready for use.