title: RNA-seq of NP550hTert, an immortalized nasopharyngeal epithelial cell line.
design: Total RNA was extracted from cell lines and xenografts using Trizol reagents following manufacturers instructions. The rRNA in total RNA was depleted by Ribo-Zero kit. After the rRNA was depleted, the remaining RNA was purified, fragmented and primed for cDNA synthesis. The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random hexamers. It was followed by second strand cDNA synthesis using DNA Polymerase I, RNase H and dUTP. These cDNA fragments then went through an end repair process, the addition of a single A base, and then ligation of the adapters. The products were then purified and enriched with PCR to create the final cDNA library. Next-generation sequencing (100 bp, paired-end) using the Illumina HiSeq 2000 sequencing system was then performed.