GSM3958068: cyto_shCtrl_rep2; Mus musculus; RNA-Seq
Polysome profiling was performed on four replicates of each condition as previously described (Gandin et al. J Vis Exp JoVE, 2014). Briefly, cytosolic lysates were loaded on a 5-50% sucrose gradient allowing for isolation of mRNAs associated with more than 3 ribosomes (hereafter referred to as polysome-associated mRNA) after ultracentrifugation. Total cytosolic RNA was isolated in parallel (Gandin et al. J Vis Exp JoVE, 2014). RNAseq libraries were prepared using the Smart-seq2 method as described previously (Picelli et al, 2014), using 10ng RNA as input material. After pre-amplification PCR, 80pg of cDNA was used for tagmentation using the Nextera XT Kit (Illumina) in a total volume of 25uL.
GEO Accession
GSM3958068