title: GSM8293910: ID8 omentum tumor - HSCDA Proficient mouse - ear tag 704; Mus musculus; RNA-Seq
library_construction_protocol: Omentum tumors were homogenized in Qiagen RLT Plus buffer supplemented with 1% 2-mercaptoethanol at 4 °C using a Biospec Products Minibeadbeater. RNA was isolated using Qiagen RNeasy Plus mini kit. Ribosomal RNA depletion was performed using QIAseq FastSelect -rRNA kit (Qiagen #335376). Step 1 of fragmentation was performed for 7 minutes at 94 °C, followed by FastSelect hybridization. First-strand synthesis, second-strand synthesis, A-tailing, adapter ligation, and library amplification were performed using the KAPA mRNA HyperPrep kit (Roche #08105952001). We used 5 μL of 1.5 μM KAPA single index adapters for Illumina (Roche #0800577001) for adapter ligation. We performed 12 amplification cycles. KAPA Pure magnetic beads (Roche #08105901001) were used for all cleanup steps