GSM7566672: EGFP(R96W)_mRNA_rep1; Homo sapiens; RNA-Seq
RNA-seq experiments were performed on 3 independent replicates from HEK293 cells overexpressing the mistranslation reporters with and without cognate mc-tRNAs. HEK293 cells were sorted on the FACSAria Fusion 5-18 cell sorter for mCherry+, i.e., successfully transfected, cell populations 48 h post-transfection. Total RNAs were extracted from sorted cells with TRIzolTM reagent (Invitrogen Cat. 15596026). 1 µg of total RNA per sample was used as input for RNA-seq library construct. Total RNA samples were polyA-selected to enrich mature mRNA species. RNA quality and quantity was assessed using the Agilent bio-analyzer. Strand-specific RNA-SEQ libraries were prepared using a TruSEQ based mRNA-SEQ library protocol (Illumina). Library quality and quantity was assessed using the Agilent bio-analyzer and libraries were sequenced using an Illumina NovaSEQ6000 (Illumina provided reagents and protocols).