GSM9099837: Hep3B, No_L2HG #1; Homo sapiens; RNA-Seq
RNA was extracted using the PureLink RNA Mini Kit (Invitrogen). 0.5 µg of total RNA was used for the constrcution of sequencing libraries. For metabolic screen, genomic DNA from HuH7 cells were extracted by using the phenol-chloroform method. For ATAC-seq, a total of 50,000 cells per condition were harvested, washed with ice-cold PBS, and lysed in ATAC lysis buffer (provided by the assay kit). Nuclei were collected by centrifugation at 500xg for 10 minutes at 4°C. For RNA-seq, libraries were prepared using the NEBNext Ultra RNA Library Prep Kit (New England Biolabs), following the manufacturer's protocol. For metabolic library screen,  sgRNA inserts were PCR-amplified using Q5 High-Fidelity DNA Polymerase (New England Biolabs, Cat# M0491) with 100 ug of gDNA per sample  following the manufacturer's protocol. For ATAC-seq, library preparation was performed using the ATAC-seq kit (Active Motif, Cat# 53150) according to the manufacturer's instructions.