strain: SD rat Sex: male treatment: pair-feeding to ZR for 16 days from 5-wk-old
Extracted molecule
total RNA
Extraction protocol
Total RNA samples were isolated from individual diencephalons using the TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and cleaned up using the RNeasy Mini Kit with a DNase treatment (Qiagen, Valencia, CA, USA). The RNA quality and quantity were examined by agarose gel electrophoresis and a spectrophotometer.
Label
biotin
Label protocol
One hundred nanograms of total RNA from each sample was reverse-transcribed, amplified and labeled using the GeneChip 3’IVT Express Kit (Affymetrix, Santa Clara, CA, USA) according to the manufacturer’s protocol.
Hybridization protocol
Labeled aRNA samples were hybridized to GeneChip Rat Genome 230 2.0 Arrays (Affymetrix). The arrays were washed, stained with the Fluidics Station (Affymetrix).
Scan protocol
Scanned with the GeneChip Scanner (Affymetrix).
Data processing
Data acquisition was performed using the GeneChip Operating Software (Affymetrix) and analyzed using the MAS5 program (Affymetrix).
