GSM3640109: A375_PRL3_B11; Homo sapiens; RNA-Seq
90-150 µg total RNA were harvested from 4sU labelled EV and PRL3 cells using Trizol reagent. The purification of 4sU labelled RNA was performed using EZ-link Biotin-HPDP at a concentration of 1 mg /ml. Following chloroform extraction, the biotinylated RNA was magnetically labelled with µMACS Streptavidin MicroBeads and purified using streptavidin µMacs columns. The eluate RNA was rRNA depleted following manufacturer's instructions. Approximately 20 ng of the rRNA depleted 4sU labelled RNA was used for the construction of cDNA sequencing libraries. cDNA libraries were prepared using NEBNext Ultra Directional RNA library Prep kit for Illumina (NEB) following manufacturer's instructions. NEBNext® Multiplex Oligos for Illumina® were used to barcode the RNA replicates. Libraries were sequenced on the HiSeq 4000 150PE following the manufacturer's protocols.
GEO Accession
GSM3640109