Fresh tumor tissue was minced, dissociated, washed and then cultured in neural stem cell media. After 5-7 days, cells or spheres were plated onto laminin-coated dishes. Cells were maintained and passaged as adherent cultures.
Extracted molecule
total RNA
Extraction protocol
RNeasy Mini kit (Qiagen)
Label
biotin
Label protocol
250 nanograms of total RNA from each sample were used to generate amplified and biotinylated sense-strand cDNA from the entire expressed genome according to the Ambion WT Expression Kit( P/N 4425209 Rev C 09/2009) and Affymetrix GeneChip® WT Terminal Labeling and Hybridization User Manual (P/N 702808 Rev. 6, Affymetrix Inc., Santa Clara, CA)
Hybridization protocol
GeneChip® ST Arrays ( GeneChip® Human Exon 1.0 ST Array) were hybridized for 16 hours in a 45°C incubator, rotated at 60 rpm. According to the GeneChip® Expression Wash, Stain and Scan Manual (PN 702731 Rev 2, Affymetrix Inc., Santa Clara, CA) the arrays were then washed and stained using the Fluidics Station 450
