title: NCHU_M&A_human  1152 cDNACHIP
status: Public on Dec 31 2008
contact.city: Taichung
contact.first: Chen-Yu
contact.last: Lee
contact.email: cu.lee@msa.hinet.net
contact.address: 250, Kuo Kuang Rd.
contact.department: Institute of Biomedical Sciences
contact.country: Taiwan
contact.institute: National Chung Hsing University
organism: Homo sapiens
manufacture_protocol: We obtained 1,152 human EST clones with putative gene names related to cell adhesion, motility, angiogenesis, signal transduction, tumorigenesis, and metastasis, etc. (Chen JJ, Peck K, Hong TM etc. Global analysis of gene expression in invasion by a lung cancer model. Cancer Res 61:5223-5230, 2001) from the IMAGE consortium libraries through its distributor (Research Genetics, Huntsville, AL). Three copies of 1,152 PCR-amplified cDNA fragments were prepared by arraying machine built in house and spotted per microarray membrane (measuring 18 X 27 mm). The whole system included tool pin, adapter, washing slots, sampling slots, sample distribution region, program and automatically changing plate system. Those treated pins were fitted into multiple-pore adapter, which is designed specially to make the pins elastic while spotting samples on membrane. The adapter could be changed for single or multiple pins system according to experimental need. The PCR-amplified cDNA in V-bottomed 96-well microtiter plates were distributed onto a positively charged nylon membrane with spots spaced 150 um apart by an arraying machine. The arraying machine was a computer controlled XYZ translation system outfitted with Teflon-coated tool pins for sample delivery. It was designed and constructed by Wittech company (Taipei, Taiwan) with position resolution and repeatability better than ± 5 um. Samples were held at the tip of the pins for delivery by the action of surface tension, and the delivery volume was governed by the diameter of the tips. Every time after DNA was spotted on membranes, by means of movement of translation stage, the solid pin was immersed into washing slots, and washed by 95% ethanol, and deionized water respectively with sonicating.