title: GSM9473901: DG_2_Dark; Mus musculus; RNA-Seq
library_name: GSM9473901
library_construction_protocol: Total RNA was extracted from approximately 20-30 mg of frozen brain tissue using the RNeasy Mini Kit, Qiagen, including the on-column DNase I digestion step according to the manufacturer's instructions. RNA integrity was verified by an Agilent 2100 Bioanalyzer, and all samples had an RNA Integrity Number (RIN) > 8.0. RNA concentration was measured using a NanoDrop spectrophotometer. Strand-specific RNA-seq libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina, NEB, following the manufacturer's protocol. Briefly, mRNA was enriched from 1 μg total RNA using poly-T oligo-attached magnetic beads, fragmented, and first-strand cDNA was synthesized. After second-strand synthesis, adaptors were ligated, and the libraries were amplified with 12-14 cycles of PCR. Final libraries were size-selected (300-400 bp) and quantified by qPCR (KAPA Library Quantification Kit) before pooling for sequencing.
identifiers.namespace: GEO
identifiers.id: GSM9473901_r1