title: GSM8028020: HC11_vangl2_knockout01_rep03; Mus musculus; RNA-Seq
library_construction_protocol: HC11 cells were harvested in 500 µL trizol and flash frozen in liquid nitrogen. Upon thawing, samples were mixed with 100 µL of 1-bromo-3-chloropropane (BCP) (Sigma, #B9673), transferred to phase lock heavy gel tubes, and stored at room temperature for 10 minutes. To separate the phases samples were centrifuged at 13,000 × g and 4°C for 15 min. The aqueous top phase was transferred to a fresh tube and mixed with 0.7x volumes of ice-cold isopropanol and 1.5 µL of GlycoBlue Coprecipitant (15 mg/mL). Samples were stored at -80C overnight followed by another centrifugation at 13,000 × g at 4°C for 30 min. RNA pellets were washed 3× with ice-cold 75% ethanol and finally resuspended in 15 µL water. NEB Ultra RNA library prep kit for Illumina