title: ONT direct RNA sequencing of MCF7 - 3 hours 100nM Estradiol induction
design: MCF7 cells were cultured in a phenol red-free starvation medium containing 10% dextran-coated charcoal-stripped FBS, 2% L-glutamine, 1% P/S, and 1% Na/Pyr for 72 hours. After hormone deprivation (HD) for 72 hours, cells were treated with 100 nM 17 -Estradiol (E2, Sigma-Aldrich, CAT#E2257) for 45 minutes, 3 hours, and 12 hours. Total RNA was isolated with TRIzol reagent (Zymo)  and poly(A) enrichment was done by Dynabeads Oligo(dT)25 (Thermo Scientific). MinION Mk1C and direct RNA sequencing protocols were used as suggested by Oxford Nanopore Technologies (ONT) with Direct RNA Sequencing Kit (SQK-RNA002) and Flow Cell (FLO-MIN106D)