GSM7103019: GS022; Homo sapiens; RNA-Seq
Tissues were snap frozen in liquid nitrogen and stored at -80*C until use. To isolate RNA, snap-frozen tissues were added to 300 mL of RNA lysis buffer (Qiagen) with a stainless-steel bead (BioSpec, 11079123) and homogenized with a Mini-Beadbeater (Biospec, C321001) for 5-10 seconds or until tissue samples were not visible. RNA was then isolated using a Qiagen RNeasy Micro Kit (Qiagen, 74004), according to manufacturer's instructions. RNA for each sample was then analyzed for quality/quantity using an Agilent bioanalyzer, Qubit 2.0 at Columbia's Molecular Pathology core. cDNA amplification for Sequencing was performed the SMART-Seq® v4 Ultra® Low Input RNA Kit (TaKaRa) for according to manufacturer's instructions. A library was  prepared using the Nextera XT DNA Library Preparation kit (Illumina) with 100-150pg starting material, ccording to manufacturer's instructions. Libraries were sequenced to a targeted depth of ~40M 100bp paired-end reads on a NovaSeq 6000