title: GSM5425796: Lung cancer tissue2; Homo sapiens; OTHER
library_construction_protocol: Total RNA of H1299 cells or the tissues were extracted and rRNA was removed. Then the clean RNA was incubated with m6A antibody for 2 hours at 4 °C. The mixture was then immunoprecipitated by incubation with Protein A beads for 2 hours at 4 °C. Captured RNA was purified and used for the following library construction according to the manufacturer's instructions. 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.