title: GSM5348271: RNA-seq.KO.PCIF1.rep3; Homo sapiens; RNA-Seq
library_construction_protocol: After the extraction of total RNA, for RNA-seq, ribosomal RNA was depleted by RiboMinus Transcriptom Isolation Kit (life technology, K1500-02). For m6A-exo-seq, mRNA was purified using Magnetic mRNA Isolation Kit (New England Biolabs, S1550S) For m6A-exo-seq, mRNA was fragmented with Fragmentation Reagents Kit (Invitrogen, AM8740) following the manufacturer's protocol, then phosphorylated with T4 PNK (NEB, M0201S), and then treated with Terminator 5'-Phosphate-Dependent Exonuclease (Lucigen) to remove phosphorylation of transcripts. Lastly, Cap-Clip (CellScript) were added to remove capped transcripts. 10% uncapped 5'RNA fragments were reserved as input, the rest of fragmented RNA were proceeded to do the immunoprecipitation with anti-N6-methyladenosine (m6A) antibody (abcam, ab151230). Immunoprecipated and input RNA for m6A-exo-seq as well as ribosomal depleted and fragmented RNA for RNA-seq were both subjected to library preparation using a TruSeq mRNA library preparation kit (Illumina) RNA-seq and m6A-exo-seq. RNA-seq was sequenced at Illumina HiSeq 4000 platform with single-end model. m6A-exo-seq was sequenced at  Illumina NovaSeq 6000 platform with paired-end model