Comment[ArrayExpressAccession] E-GEOD-46809 MAGE-TAB Version 1.1 Public Release Date 2013-10-01 Investigation Title The gene expression on human hair root Comment[Submitted Name] The gene expression on human hair root Experiment Description In HAIR experiment of astronauts, the hair samples are limited to only five strands from each astronaut. Therefore, it is essential to develop the method to analyze no less than five hair roots. To achieve this purpose, in this study, we performed the microarray analyses using the extracted RNA from one, five, or ten hair roots. 13 samples from two healthy subjects Term Source Name ArrayExpress EFO Term Source File http://www.ebi.ac.uk/arrayexpress/ http://www.ebi.ac.uk/efo/efo.owl Person Last Name Seki Terada Seki Person First Name Masaya Masahiro Masaya Person Email m_seki@aes.co.jp Person Affiliation Kagoshima Univ. Person Address Graduate School of Medical and Dental Sciences, Kagoshima Univ., Sengen 2-1-1, Tsukuba, Ibaraki, Japan Person Roles submitter Protocol Name P-GSE46809-1 P-GSE46809-3 P-GSE46809-4 P-GSE46809-2 P-GSE46809-5 Protocol Description The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded. ID_REF = VALUE = Normalized signal intensity Cyanine-3 (Cy3) labeled cRNA was prepared from 0.1 ug RNA using the Amino Allyl MessageAmp II aRNA Amplification kit (Ambion) according to the manufacturer's instructions, followed by RNeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer. 600 ng of Cy3-labelled cRNA was fragmented at 60M-0C for 30 minutes in a reaction volume of 0.025 ml containing 25x fragmentation buffer and 10x blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 0.025 ml of 2x GEx Hybridization Buffer HI-RPM was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4851A) for 17 hours at 65M-0C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37M-0C GE Wash buffer 2 (Agilent) each GE Wash Buffers were all supplied with 0.005% Triton X-102, then dried immediately by brief centrifugation. Approximately 2-3 mm of the hair root was used as sources for RNA extraction. The hair root was cut into about 15 fragments (each fragment is 0.1-0.2 mm) using micro surgical knife under stereoscopic microscope. The collected fragments were immersed in 800 M-NM-