title: GSM6638936: IRF1KO, Control, RepC; Homo sapiens; RNA-Seq
library_construction_protocol: Samples were lysed, and RNA was extracted using QIAGEN RNeasy mini kit per manufacturer's instructions. Total RNA was then submitted to Genome Technology Access Center (GTAC), Washington University in St. Louis, School of Medicine for Next generation mRNA sequencing. Briefly, library preparation was performed using ~ 1ug of total RNA processed using RiboErase kits to remove ribosomal RNA (Kapa Biosystems). mRNA was then fragmented in reverse transcriptase buffer and heating to 94 degree Celsius for 8 min. mRNA was reverse transcribed to yield cDNA using SuperScript III RT enzyme (Life Technologies, per manufacturer's instructions) using random hexamers. A second strand reaction was performed to yield ds-cDNA that was blunt ended, extended by adding an A base to the 3' ends, and followed by ligation of Illumina sequencing adapters. Ligated fragments were then amplified for 12-15 cycles using primers incorporating unique dual index tags. Fragments were sequenced on an Illumina NovaSeq-6000 to generate paired end reads extending 150 bases.