title: GSM6283568: Panc1 WT cells; Homo sapiens; RNA-Seq
library_construction_protocol: Remove medium, wash the cells twice with PBS and detach by adding trypsin-EDTA and incubating at 37 °C for 3 min. Add culture medium and harvest 106 cells.  Spin down samples at 300 x g for 3 min, remove supernatant and extract total mRNA using commercially available silica membrane spin columns, by following the manufacturer`s protocol.  Determine the concentration and purity of total RNA by using a UV-Vis spectrophotometer. The samples were stored at - 80 °C.  Total RNA integrity was assessed by electrophoresis of 200 ng of sample on 1% agarose gel stained with ethidium bromide. Use 2 µg 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 a bioanalyzer system. Pool the libraries in a single sample up to a final concentration of 10 nmol/L and measure it with the bioanalyzer. Use high-throughput sequencing system with Single Read 100 bp mode.