GSM5693386: PancreaticTumour_Pkn2Wt_B; Mus musculus; RNA-Seq
Tumours were extracted and stored in RNAlater (ThermoFisher Scientific, AM7020) at -20oC. To extract RNA, samples were macerated using the TissueLyzer II (QIAGEN) with 5 mm TissueLyzer beads (QIAGEN, 69989) in RLT buffer. RNA was prepared using the RNeasy mini kit (QIAGEN) as per manufacturer's instructions. RNA samples were quantified using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked with Agilent Fragment Analyzer (Agilent Technologies, Palo Alto, CA, USA) The library was prepared using NEB Next Ultra II Directional RNA Library Prep Kit for Illumina using manufacturer's recommendations (NEB).