title: GSM6865253: normal3, MeRIP-seq; Homo sapiens; RIP-Seq
library_construction_protocol: Total RNA extraction was provided by Cloud-seq Biotech Inc. (Shanghai, China). RNA was extracted using TRIzol Reagent (Invitrogen, Carlsbad,CA, USA) according to the manufacturer's instructions.A NanoDrop ND-1000 instrument (Thermo Fisher Scientific, Waltham, MA, USA) was used to measure the RNA concentration of each sample, controlling OD 260/280 values of 1.8-2.1 to ensure good RNA purity. The m7G-IP-Seq service was provided by CloudSeq Inc. (Shanghai, China). Total RNA was decapping with mRNA decapping enzyme (New England Biolabs), and then subjected to immunoprecipitation with the GenSeq® m7G-IP Kit (GenSeq Inc.) by following the manufacturer's instructions. Briefly, RNA was randomly fragmented to ~200 nt with RNA fragmentation reagents supplied in the kit. Protein A/G beads were coupled to the m7G antibody by rotating at room temperature for 1 h. The RNA fragments were incubated with the bead-linked antibodies and rotated at 4°C for 4 h. After incubation, the RNA/antibody complexes were washed for several times, and then, captured RNA was eluted from the complexes and purified. RNA libraries for IP and input samples were then constructed with GenSeq® Low Input Whole RNA Library Prep Kit (GenSeq, Inc.) by following the manufacturer's instructions. Libraries were qualified using Agilent 2100 bioanalyzer and then sequenced in a NovaSeq platform (Illumina). MeRIP-seq