title: GSM4775599: Tu8902_shGOT1_plsdox_2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using the RNeasy Mini Kit (Qiagen, 74104) and reverse transcription was performed from 2 µg of total RNA using the iScript cDNA synthesis kit (BioRad,1708890) according to the manufacturer's instructions. Samples (RIN > 7) were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (cat. E7760L) with Poly(A) mRNA Magnetic Isolation Module (cat. E7490L) and NEBNext Multiplex Oligos for Illumina (unique dual-index primers; cat. E6440L). PolyA+ mRNA was fragmented (10 min) and reverse-transcribed into first-strand cDNA, followed by end repair, dA tailing, and ligation of NEBNext adapters. Libraries were amplified using 11 PCR cycles and assessed by TapeStation (Agilent) and qPCR using Kapa's library quantification kit for Illumina Sequencing platforms (cat. KK4835).