title: RNA-seq of Homo sapiens: ventricular cardiomyocyte differentiation from WTC11 at D6 rep1
design: Cells were grown in 1 well of a 6- or 12-well dish per replicate. Cells were dissociated in 0.25% Trypsin-EDTA for 10 mins, followed by quenching in 15%FBS in DPBS. Cells were lifted off the dish and resuspended to single cells by repeatedly using P1000 pipette and then transferred to a 1.5 ml tube and centrifuged at 300 x g for 5 min at room temperature. Pellets were snap frozen in dry ice. Total RNA was isolated using Qiagen RNeasy micro kit (Qiagen #74004) with QIAshredder (Qiagen #79656) and eluted in 25 l water. RNA concentration was determined using Nanodrop and quality was checked using Agilent Bioanalyzer. RNA-seq libraries were generated using Illumina Stranded Total RNA Prep with Ribo-Zero Plus kit were prepared as per manufacturers instructions using 400 ng of input RNA. Library concentration was quantified using Qubit Fluorometer and dsDNA HS Assay Kit and quality assessed on Agilent Bioanalyzer.