title: GSM8891601: QGP1_sgPIKFYVE_1_Rep2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from cells using miRNeasy Kit (QIAGEN) following the manufacturer's instructions after cells were lysed in QIAzol Lysis Reagent. RNA concentration and purity were assessed using NanoDrop. RNA integrity was assessed using the Agilent Bioanalyzer with the Eukaryote Total RNA Nano Kit (Agilent Technologies, #5067-1511). Ribosomal RNA was removed enzymatically using the KAPA RNA HyperPrep Kit with RiboErase HMR (Roche; catalog #08098140702), followed by heat fragmentation of RNA to ~200–300 bp in fragmentation buffer. First-strand cDNA synthesis was performed using reverse transcriptase and random primers, and double-stranded cDNA was generated through second-strand synthesis. End-repair and adapter ligation were carried out using New England Biolabs (NEB) adapters, and libraries were amplified with KAPA HiFi HotStart mix and NEB dual barcodes.