Illumina HiSeq 1500 sequencing; Large-scale study of total and polysomial mRNA after DDX6 depletion in HEK293 cells
Large-scale study of total and polysomial mRNA after DDX6 depletion in HEK293 cells
Human embryonic kidney HEK293 cells  were maintained in DMEM supplemented with 10% (v/v) fetal calf serum. siRNA transfections were performed using Lipofectamin 2000 (Life Technologies, France) according to the manufacturer's protocol. Cells were harvested 48h post-transfection Total and polysomial RNA was extracted using the SV Total RNA Isolation System (Promega) according to the manufacturers instructions Library preparation was performed at the Ecole Normale Superieure Genomic Platform (Paris, France). Ribosomal RNA was depleted from 1 g of total or polysomial RNA using the Ribo-Zero kit Human/Mouse/Rat (Epicentre). Libraries were prepared using the strand specific RNASeq library preparation ScriptSeq V2 kit (Epicentre), and multiplexed by three on flow cell lanes.