GSM9312181: hESC, FUBP3, rep2 (second run); Homo sapiens; RNA-Seq
Sanger sequencing was used to conﬁrm proper insertion and then control and RBP-ADAR plasmids were infected into hESCs by transfection reagent jetOPTIMUS (Polyplus, catalog#101000051). After transient transfection into target cells for 24 h, the cells containing RBP-ADAR fusion proteins or mCherry-ADAR proteins were sorted by GFP and mCherry ﬂuorescence and using TRIzol reagent to extract the mRNA from the sorted cells. mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. After library quality control, quantified libraries were pooled and sequenced on Illumina NovaSeq 6000 with paired-end 150-bp reads, providing sequencing data at 40 Gb per library.