title: GSM6865217: shMETTL3; Homo sapiens; RNA-Seq
library_construction_protocol: The total RNA was extracted from both control and shMETTL3-1 CD4+ T cells using RNeasy Plus Mini Kit (Qiagen) and the mRNA purification was further achieved using PolyATtract mRNA Isolation System (Promega). The mRNA concentration was measured by Qubit RNA HS assay kit (ThermoFisher) and the quality was confirmed by Agilent Bioanalyzer RNA Pico assay chip. Then, the mRNA was subjected to library construction using Oxford Nanopore direct RNA sequencing kit (SQK-RNA002) according to the manufacturer's manual. A total of 500 ng mRNA was first annealed and ligated to RT adapter and then reverse-transcribed to form DNA-RNA hybrid products. Then, RNA adapter was ligated to the products and was ready for sequencer loading. The resulting library was sequenced on R9.4.1 flow cell (FLO-MIN106D) using a MinION sequencer.