title: GSM8196500: 5xFADEif2ATRAP_rep3; Mus musculus; RNA-Seq
library_construction_protocol: The brain tissue was thawed in an ice-cold  glass homogenizer containing 1 mL cell-lysis buffer. After manually homogenizing the samples with 3-5 gentle strokes, samples were  homogenized in a motor-driven overhead stirrer. A post-nuclear supernatant was prepared by centrifugation. Post-mitochondrial supernatant was prepared by centrifugation after NP40 and DHPC treatment. GFP-antibod-coated beads were added to each supernatant. After overnight incubation, the polysome-bound beads were washed with the high-salt buffer. RNA clean-up was performed using RNeasy Mini Kit (Qiagen) following the manufacturer's instructions. libraries were constructed using SMART-Seq® mRNA LP (Takara, 634768), multiplexed using Unique Dual Index Kit 1–96 (Takara, 634752), and size-selected using NucleoMag® NGS Clean-up and Size Select beads (Takara 744970.5) as per the manufacturer's recommendations.