title: NTU_CGM_MCF Human 672 Metachip
status: Public on May 12 2006
contact.city: Taipei
contact.first: Sung-Liang
contact.last: Yu
contact.email: slyu@ntu.edu.tw
contact.address: Jen Ai Road Section1
contact.department: Clinical Laboratory Sciences and Medical Biotechnology
contact.country: Taiwan
contact.institute: National Taiwan University
contact.phone: 886-2-23958341
organism: Homo sapiens
manufacture_protocol: 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 mm 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 mm. 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.
  The nylon membranes carrying 672 PCR-amplified cDNA fragments were prepared by arraying machine built in house. The 672 invasion-associated genes identified in our previous study were re-arrayed in duplicate on a nylon membrane by arraying machine built in house (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).