title: NTU_CGM_MCF Mouse 6.1k Microarray
status: Public on May 24 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: Mus musculus
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 (measured 18 mm by 27 mm) carrying 6,144 PCR-amplified cDNA fragments were prepared by arraying machine built in house. The cDNA microarray membrane contains 6,144 non-redundant expressed sequence tag (EST) clones with putative gene names selected from the IMAGE consortium mouse cDNA libraries based on the insert length and sequence of the EST clones in the Unigene database.