title: GSM5509540: lncRNA Capture-Seq RC rep3; Mus musculus; OTHER
library_construction_protocol: lncRNAcapseq: ILPFC tissue was collected immediately after RC or EXT training (N=24). 2mg of total RNA isolated from ILPFC tissue (4 pooled per library, 3 libraries per group) was used to generate the RNA library. Total RNA was treated with NEBNext® rRNA Depletion Kit (NEB) to remove rRNA. ATAC-seq: Five ILPFCs per sample were pooled together for FACS. After FACS, Arc+NeuN+ populations from RC or EXT group (n=3 libraries per pooled group) were used for ATAC-seq in following procedure. 50,000 cells were resuspended in 50 μl 2x lysis buffer (20 mM Tris–Cl, pH 7.4; 20 mM NaCl; 6 mM MgCl2; 0.10% Igepal CA-630) and spun down immediately (500 g for 10 min at 4 °C). RNA libraries were then generated by NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (NEB). A custom-designed probe panel (Roche), which targets 28,228 known and predicted mouse lncRNAs (Bussotti et al, 2016), was used to capture lncRNAs in accordance with the manual of SeqCap EZ Hybridization and Wash Kit and SeqCap EZ Accessory Kit (Roche). Captured lncRNA libraries were sequenced on an Illumina HiSeq 4000 platform with the read length of 150 bp x 2. For ATAC-seq, the transposase reaction was performed using the ATAC-seq kit (Active motif) following the manufacturers protocol. Paired-end (PE) libraries were sequenced using an Illumina HiSeq 4000 sequencing platform with the read length of 150 bp x 2.