title: GSM4222603: WT_IP; Homo sapiens; RIP-Seq
library_construction_protocol: Total RNA was extracted using Trizol reagent (Invitrogen, CA, USA) following the manufacturer's procedure. mRNA was purified using poly-T oligo attached magnetic beads (Invitrogen). mRNA was then fragmented into ~100nt oligonucleotides using divalent cations under elevated temperature. Fragmented mRNA was incubated with m6A-specific antibody in IP buffer (50 mM Tris-HCl, 750 mM NaCl and 0.5% Igepal CA-630) supplemented with 0.5 μg/μl BSA for 2h at 4 ℃. Immunoprecipitated m6A RNA fragments were used for the construction of m6A-seq libraries (IP). And 1 ug of total RNA was used for the construction of RNA-sequencing libraries (Input). RNA libraries were prepared for sequencing using standard Illumina protocols. Immunoprecipitated m6A RNA fragments for m6A-seq (IP), total RNA for RNA-Seq (Input).
attributes.tag: GEO Accession
attributes.value: GSM4222603