GSM8346378: Tumor, HS3ST1 KO, rep 2; Mus musculus; RNA-Seq
Tumor samples were incubated with TRIzol (Thermo Fisher) and homogenized using the TissueLyser II system (Qiagen). Samples were spun at >10,000 rpm to remove cell debris. Samples were incubated with 200 µL chloroform per 1mL Trizol for 2-3 min at room temperature. To separate the aqueous phase, samples were spun at >10,000rpm for 18min at 4C. The aqueous phase was separated into a new tube and an equal part of 70% Ethanol was added. RNA was isolation using the RNEasy kit (Qiagen) as described by the manufacturer with 1 extra Buffer RPE wash. RNA integrity was verified on a Tapestation (Agilent). RNA libraries were prepared by the UCSD IGM sequencing core using the Illumina Total RNA Prep with Ribodepletion kit