Biotinylated cRNA was prepared from the total RNA according to the Affymetrix protocol (Expression Analysis Technical Manual Rev. 2, 2005-2006, Affymetrix)
Hybridization protocol
Following fragmentation, cRNA was hybridized on the GeneChip Array and the GeneChips were washed and stained in the Affymetrix Fluidics Station 450 according to the standard Affymetrix protocol (Expression Analysis Technical Manual Rev. 2, 2005-2006, Affymetrix).
Scan protocol
GeneChips were scanned using the Affymetrix GeneChip 3000 7G scanner.
Description
This sample was analyzed as part of the Stem Cell Genomics Project (http://www.stembase.ca). The biological material was submitted to the Ontario Genomics Innovation Centre (http://www.ottawagenomecenter.ca/) by Dr. Michael Rudnicki (mrudnicki@ohri.ca; 501 Smyth Road) for analysis. Stembase Experiment ID: E233 Stembase Experiment ID link: http://www.stembase.ca/?path=/browse/experiment&id=233 SCGP Sample ID: S333 SCGP Sample ID link: http://www.stembase.ca/?path=/browse/experiment&id=233#SAMPLE_342 Short description: Time Course in vitro Differentiation of Myogenic Primary Myoblast into Myotubes for Ontario Genome Project 2004-05. , The Differentiation is leaded by removing the Proliferation Medium (Ham's F10 Medium + growth factors) and feeding with Differentiation Medium (DMEM hg + 2.5% Horse serum). , The expected data are: % Viability of cells; Total RNA extraction; BioAnalysis and MicroArray of Undifferentiated and Differentiated cells., Time course: 0 hr Estimated purity: 100% RNA concentration: 1170.2 ng/µL Num cells for RNA prep: More than 2 x 10E+6 Balb/c Sample volume: 27µL
Data processing
Calculated using the MAS5 algorithm where sc=1500, tau=0.015, alpha1=0.04, and alpha2=0.06