genotype: L26 daDREAM transgenic line tissue: Hippocampus
Growth protocol
Experiments were performed in adult male mice from two daDREAM transgenic lines, L1 and L26, homozygous for the transgene, and wild type littermates. The genetic background in all cases was C57B6xCBA.
Extracted molecule
total RNA
Extraction protocol
RNA was prepared using TRIzol (Invitrogen) and the RNeasy Mini Kit (Qiagen). RNA was quantified and the RNA quality was assessed with a 2100 Bioanalyzer from Agilent Technologies
Label
biotin
Label protocol
Biotinylated cRNA was prepared according to the standard Affymetrix protocol from 4 µg total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
Hybridization protocol
Following fragmentation, 10 µg of biotinylated cRNA were hybridized to Affymetrix chips (GeneChip Mouse Genome 430 2.0 Array). Each sample was added to a hybridization solution containing 100mM 2-(N-morpholino)ethanesulfonic acid, 1 M Na+, and 20mM of EDT
Scan protocol
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G
Description
Gene expression data from hippocampus cells (L26)
Data processing
GeneChip intensities were background-corrected, normalized and sumarized by RMA method, using the “Affy” package from Bioconductor.