GSM8254560: GN035_+iM0 hECT; Homo sapiens; RNA-Seq
Tissues were snap frozen in liquid nitrogen and stored at -80 ºC until use. To isolate RNA, snap-frozen tissues were added to 300 µL of RNA lysis buffer with a stainless-steel bead (BioSpec, cat. no. 11079123) and homogenized with a Mini-Beadbeater (Biospec, cat. no. C321001) for two cycles of10 second intervals, with storage on ice between cycles. A Qiagen RNeasy Micro Kit (Qiagen, cat. no. 74004) was then used according to the manufacturer's instructions. Total RNA for each sample was quantifiedusing an Agilent 2100 Bioanalyzer with an RNA Pico Chip at Columbia's Molecular Pathology core. RNA samples with an RNA integrity number (RIN) > 8 were considered acceptable for further analysis. A library was prepared through Columbia's Genome Center using a Nextera XT DNA Library Preparation kit (Illumina TruSeq Chemistry) with 100-150pg starting material, following the manufacturer's instructions.