title: RNA-seq with metabolic labelling of human breast cancer cell line MCF7 treated with a panel of 40 different cancer drugs and untreated controls
design: RNA-seq with metabolic labelling of human breast cancer cell line MCF7 treated with a panel of 40 different cancer drugs and untreated controls
library_construction_protocol: MCF-7 cells were cultured at 50000 cells per well in 96-well plate overnight. Cells were treated for four hours with compounds from a panel of cancer drugs at a concentration of 10, 100, 1000, 10000 or 100000 micromolar for each compound in DMSO, or with only DMSO. Additionally, 200 micromolar 4sU was added four hours before cell harvest. RNA was isolated using magnetic beads following the BOMB-bio protocol 8.2 version 1.0 The sequencing libraries were prepared from isolated RNA through mRNA capture with an oligo-dT primer, and full length cDNA molecules were constructed through the use of a template switching oligo. The resulting cDNA fragments were pre-amplified through PCR and then fragmented through tagmentation to yield sequence ready fragments.