title: GSM7210759: Sup. Fig. 6, HeLa-R634Q ICS screen, input, lib 2; Homo sapiens; OTHER
library_name: GSM7210759
library_construction_protocol: RNA from HeLa: Direct-Zol (Zymo) kit was used according to manufacturer's instructions. DNA from HeLa: cells were palleted and genomic DNA was extracted using NEB Monarch genomic DNA purification kit, including the RNase treatment and elution in 50 µl elution buffer; 100-500 ng of DNA was used for library preparation. HeLa RNAseq: Library preparation for HeLa cells was done based on RNA extracts from live cells. Barcoded stranded mRNA-seq libraries were prepared from 150ng of high quality total RNA samples using the NEBNext Poly(A) mRNA Magnetic Isolation Module and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs (NEB), Ipswich, MA, USA) implemented on the liquid handling robot Beckman i7. Obtained libraries that passed the QC step were pooled in equimolar amounts; 2.1 pM solution of this pool was loaded on the Illumina NextSeq 500and sequenced bi-directionally, generating ~500 million paired-end reads, each 75 bases long.HeLa ICS screen: see detailed methods in Schraivogel et al., 2022.