direct RNA-seq of yeast S288C
Yeast RNA was isolated from S. cerevisiae strain S288C grown overnight at 30C in 15 mL medium containing 10 g/L yeast extract, 20 g/L peptone, and 20 g/L glucose. RNA was extracted using the ZymoBIOMICS Quick-RNA Fungal/Bacterial kit (Zymo Research, Irvine, CA, USA). A total of 600 ng total yeast RNA was used to prepare librariy. The dRNA library was produced using the SQK-RNA001 kit (ONT, Oxford, UK) which includes an RT step. After this, the motor protein was attached to the RNA strands specifically. The library was loaded onto a flow cell for a 48 hour sequencing run lasting. Direct sequencing was performed on a single R9.5/FLO-MIN107 flow cell.