GSM3701966: EV Rep 2; Homo sapiens; RNA-Seq
Cytosol is removed using a low salt buffer containing 0.03% digitonin.  Cells are washed with the same buffer containing 0.004% digitonin before the remaining membrane fraction is lysed with a 1% NP-40 high-salt buffer.  Polysomes are enriched by collecting the void of an Sephacryl-400 column and biotin labeled polysomes isolated by incubating on streptavidin beads.  The final bead-bound polysomes are extracted using an RNeasy Kit from Qiagen. Bioanalyzer is used to calculate the concentration of total RNA and 10 ng per sample is collected and libraries made using Ultra II RNA Library Kit (NEB, E7645S)
GEO Accession
GSM3701966