GSM6602220: HCT116 sh_NC_2_Input; Homo sapiens; RIP-Seq
RNA was harvested using Rneasy mini plus kit (Qiagen). 1.3 ug of total RNA was used for the construction of sequencing libraries. RNA libraries for MeRIP-seq were prepared using NovaSeq mRNA-Seq Library Prep Kit following manufacturer's protocols. Chemically fragmented RNA (100 nucleotides) was incubated with m6A antibody for immunoprecipitation according to the standard protocol of the Magna methylated RNA immune-precipitation (MeRIP) m6A Kit (Merck Millipore). Enrichment of m6A containing mRNA was then analyzed either through quantitative reverse-transcription polymerase chain reaction (qRTPCR) or by high-throughput sequencing. For highthroughput sequencing, purified RNA fragments from m6A-MeRIP were used for library construction with the NEBNext Ultra RNA library Prep kit from Illumina and were sequenced with Illumina HiSeq 2000.