GSM5002167: C2_HS-HSATIIIKD-IP-rep2; Homo sapiens; RIP-Seq
Total RNAs were prepared using TRI Reagent (Molecular Research Center, Inc.), according to the manufacturer's manual. The mRNAs were isolated Seq-Star poly(A) mRNA Isolation Kit (Arraystar) and chemically fragmented to around 100-nt sizes. m6A RIP was performed to enrich m6A methylated mRNA fragments with anti-N6-methyadenosine (m6A) antibody (Synaptic Systems, 202003). The RNA-seq libraries for both the m6A immunoprecipitated RNAs and the input RNAs were prepared with a KAPA Stranded mRNA-seq Kit. The barcoded libraries were denatured to single-stranded DNAs, captured on Illumina flow cells, amplified in situ as sequencing clusters, and sequenced for 300 cycles on Illumina HiSeq 4000 system per the manufacturer's instructions.