title: GSM6652340: SNGM_Paclitaxel_2; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted with Qiagen RNeasy Plus Columns Ribosomal RNA depletion was performed using QIAseq FastSelect -rRNA kit (Qiagen, Cat. #335376). 350 ng input RNA was used for ISH cells; 500 ng input RNA was used for SNGM, HEC265, and S33Y cells. 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, Cat. #08105952001). We used 3.33 μL of 1.5 μM KAPA single index adapters for Illumina (Roche, Cat #0800577001) for adapter ligation. We performed 11 amplification cycles. KAPA Pure magnetic beads (Roche, Cat. #08105901001) were used for all cleanup steps. Library quality was confirmed by TapeStation by the University of Colorado Cancer Center Pathology Shared Resource. Sequencing was performed on a NovaSeq6000 instrument by the University of Colorado Genomics and Microarray Core.