P1-P3 mouse pups transgenic for SOD1G93A, SOD1WT or non-transgenic pups were sacrificed by using an approved method of euthanasia. Under a dissection microscope, the parenquima were isolated and the meninges were carefully stripped away with fine forceps. The tissue was then dissected into small pieces and transferred to a solution containing 12ml of HBSS, 1.5 ml of trypsin (GIBCO) and 1% DNAse (Sigma) and incubated at 37ºC for 15 min, swirling the mixture periodically. The supernatant containing the dissociated cells was collected and 3ml of serum was added to inhibit the trypsin. Cells were then centrifuged at 1000 rpm for 5 min, resuspended in Glia medium: (Minimun Essential Medium with Earl’s salts (GIBCO), D-(+)-Glucose 20% (Sigma), Penicillin-streptomycin (GIBCO), 10% Horse Serum (GIBCO)) and plated at the concentration of 80,000 cells per mL in T75 flasks (Falcon).
Label
biotin
Label protocol
Standard procedure outlined by the Illumina TotalPrep RNA Amplification Kit.
Hybridization protocol
Standard procedure outlined by the Illumina TotalPrep RNA Amplification Kit.
Scan protocol
Arrays were read by Illumina Bead Array Reader. Analysis was done using the Illumina Bead Studio Program.
Description
B6SJL-Tg(SOD1*G93A)2Gur/J
Data processing
Average' Normalization setting was used in BeadStudio