For neural differentiations, NPCs were dissociated with Accutase and plated in neural differentiation media (DMEM/F12-Glutamax, 1XN2, 1X B27-RA, 20 ng/ml BDNF, 20 ng/ml GDNF (Peprotech), 1 mm dibutyrl-cyclicAMP (Sigma), 200 nm ascorbic acid (Sigma) onto PORN/Laminin-coated plates. 200,000 cells/well of a 6-well plate. hiPSC derived-neurons were differentiated for 6 weeks.
Extracted molecule
total RNA
Extraction protocol
Cells were lysed in RNA BEE (Tel-test, Inc). RNA was chloroform extracted, pelleted with isopropanol, washed with 70% ethanol and resuspended in water. RNA was treated with RQ1 RNAse-free DNAse (Promega) for 30 minutes at 37C and then the reaction was inactivated by incubation with EGTA Stop buffer at 65C for 10 minutes.
Label
biotin
Label protocol
GeneChip WT Sense Target Labeling and Control Reagents (900652) and GeneChip WT cDNA Synthesis and Amplification Kits (900672 and 900673)
Hybridization protocol
As recommended by Affymetrix.
Scan protocol
GeneChip scanner 3000 7G
Data processing
Data in the sample data tables are rma background corrected, quantile normalized, log2 transformed and mean probeset summarized. Further processed data on the Series record were performed by Partek Genomics Suite.
