title: GSM5425805: H1299-METTL14 RIP; Homo sapiens; RIP-Seq
library_construction_protocol: The cell lysates of H1299 were incubated with METTL14 antibody. Associated RNA-protein complexes were collected and washed, then subjected to proteinase K digestion and RNA extraction by TRIzol. Total RNA was isolated from H1299 cells.The quantity and quality of RNA were assessed by the NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific). RNA integrity was determined by RNA integrity number with the 2200 TapeStation analyzer (Agilent Technologies), and samples that had an RNA integrity number above 7.0 were considered acceptable. The isolated RNAs were used for preparing cDNA libraries by NEBNext Ultra RNA Library Prep Kit for Illumina (ENB). Products were purified with Agencourt AMPure XP beads (Beckman) and diluted to 10 pM for cluster generation in situ on a HiSeq 3000 paired‐end flow cell. The clustering of the index‐coded samples was carried out using a HiSeq Rapid PE Cluster Kit V2 (Illumina). After cluster generation, the libraries were sequenced (2 × 150 bp) on the HiSeq 3000 platform.