title: GSM7114108: Sample 8_Patu-T resistant to paclitaxel cells - biol rep 2; Homo sapiens; RNA-Seq
library_construction_protocol: Remove medium, wash the cells twice with PBS and detach by adding trypsin-EDTA and incubate at 37 °C for 5 min. Add culture medium and harvest 1-3 x 106 cells. Spin down samples at 1200 RPM for 5 min, remove supernatant and extract total mRNA using mirVana™ miRNA Isolation Kit and following the manufacturer`s protocol. Determine the concentration of total RNA by using Qubit RNA BR Assay Kit and its integrity using Agilent 2200 TapeStation system and RNA ScreenTape kit. The samples were stored at - 80 °C. Use 500 ng of total mRNA per each sample. Follow mRNA library preparation protocol according to manufacturer's instructions. Determine quality and concentration of each library by using Agilent 2200 TapeStation system. Pool the libraries in a single sample up to a final concentration of 10 nmol/L and measure it with Agilent 2200 TapeStation system. Libraries were paired-end sequenced (2x75bp) on a NextSeq500 (Illumina) according to the corresponding protocol.