GSM9073163: 10387Primary231012; Homo sapiens; RNA-Seq
For RNA-seq of primary tissue, GBO and rGBO samples were snap-frozen on dry ice and stored at -80°C until processing. The samples were homogenized in TRIzol using a disposable pestle and handheld motor. RNA purification was performed with a Zymo RNA Clean & Concentrator kit after phase separation following the manufacturer's instructions. RNA concentration and quality were measured using a Nanodrop 2000. RNA sequencing libraries were prepared using the NEBNext® Poly(A) mRNA Magnetic Isolation Module to extract mRNA, followed by library preparation with the NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® following the manufacturer's instructions. Library size, concentration and quality were assessed using the Qubit dsDNA assay and Bioanalyzer. Sequencing of samples was performed on either a NovaSeq 6000 (Illumina) or a NextSeq 550 (Illumina) with a final sequencing depth of approximately 5-10 million reads per sample.