GSM8269152: NCI-H441 shNKX2-1(6) Set2 RNA-seq Rep1; Homo sapiens; RNA-Seq
RNA was extracted from cell pellets stored at -80°C using an RNeasy Mini Kit (Qiagen) with on-column DNase digestion using RNase-Free DNase Set (Qiagen). RNA was eluted in RNase-free ddH2O, and quantified by nanodrop. RNA integrity was determined using the Agilent RNA 6000 Nano Kit on an Agilent 2100 Electrophoresis Bioanalyzer, all RNA samples used for RNA-seq had a RNA integrity number (RIN) > 9.0. 1000-2000µg of RNA was polyA-selected using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) and libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit (NEB). Final libraries were quantified using Qubit, pooled, and sequenced by Broad Walk-Up Sequencing on a Nextseq 500 using 2x40bp sequencing.