Dataset features


Application: Gene expression microarray analysis
Number of samples: 8
Release date: Nov 12 2010
Last update date: Mar 22 2012
Access: Public
Genes: AtRbcX1, SSBP3, LDB1, FXYD1, PKN2
Taxon: Caenorhabditis elegans, Homo sapiens
Dataset link Gene expression regulation by C. elegans SAM-10

Experimental Protocol

Animals were grown on fresh 8P plates ( until gravid. Eggs were collected by bleaching and hatched in M9 buffer overnight. L1 animals were fed on fresh food for 2 hours at room temperature, harvested and separated from worm debris using a 25μm mesh. RNA was isolated using a standard Trizol protocol. RNA preps were stored at -80C. Quality of RNA preps was first estimated using Agilent 2100 Bioanalyzer. RNA was then reverse-transcribed using the Genisphere Array 350 kit, which generate tagged cDNA. Resulted products are hybridized to the microarrays. Four independent high quality RNA preps (RIN>=7) were chosen for microarray assay using long oligomer-based spotted microarray (Washington University, St. Louis, MO). For dye-swap controls, we ran a technical replicate hybridization for each pair of samples in which the dyes are reversed. Gene spots with “found/good” signals (defined by ScanArray, channel flag 3) in all scans were chosen for gene expression analysis.










Michael Nonet