GSM5048749: NAT Rep4; Homo sapiens; RNA-Seq
Total RNA was isolated using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. The purity of extracted RNA was measured using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Inc., Wilmington, DE) and the concentration was assessed with Qubit 2.0 Fluorometer (Agilent Technologies, Santa Clara, CA) using an RNA HS Assay Kit. Samples with RNA Integrity Number (RIN) greater than 4 as assessed by a BioAnalyzer (Agilent) with no visible sign of genomic DNA contamination from the HS Nanochip tracings were used for total RNA library generation. RNA-seq libraries were prepared using the Illumina TruSeq Stranded Total RNA Sample Prep Kit with Ribo-Zero Gold (catalog #RS-122-2201) according to the manufacturer protocol. The sequencing was performed in paired end manner, generating 2 X 100 bp paired-end reads using the Illumina HiSeq 2500 system.  Pre-alignment data QC were assessed with FastQC. Post-alignment data quality was assessed with an in-house quality control pipeline/database for RNA-seq database (14). RNA-seq data were trimmed for any adapter sequences using Adapter Removal (15).