title: GSM4996249: PIM3_KO; Homo sapiens; RNA-Seq
library_construction_protocol: Total cellular RNA was extracted using the RNeasy kit (Qiagen) according to the manufacturer's protocol. The quality of the total RNA was assessed using the Agilent 2100 Bioanalyzer followed by 2 rounds of poly A+ selection and conversion to cDNA. The NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® library generation kit (New England Biolabs) was used per the manufacturer's instructions. The libraries were quantitated using qPCR in a Roche LightCycler 480 with the Kapa Biosystems kit for library quantitation (Kapa Biosystems, Woburn, MA). Subsequently, sequencing was performed on the Illumina NextSeq500 using the latest versions of the sequencing reagents and flow cells with single-end 75 bp reads.