title: GSM9673180: 25624-5-Lefthemisphere; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated using the RNeasy Plus Micro Kit (Qiagen), following the manufacturer's protocol. RNA concentration was measured using a Qubit 2.0 Fluorometer (ThermoFisher Scientific), and RNA quality was assessed using the Agilent TapeStation. Sequencing libraries were constructed using the NEBNext Ultra II RNA Library Prep Kit for Illumina, in combination with the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs). In brief, polyadenylated RNA was captured using oligo(dT) beads and fragmented at 94°C for 15 min. First- and second-strand cDNA synthesis followed, after which the cDNA fragments were end-repaired, A-tailed, and ligated to universal adapters. Libraries were indexed and amplified by limited-cycle PCR. Library quality was verified on the Agilent TapeStation and quantified using both the Qubit 2.0 Fluorometer and qPCR (KAPA Biosystems). Libraries were clustered on a single flowcell lane and sequenced on an Illumina HiSeq 4000 (or equivalent) platform using 2 x 150 bp paired-end reads.