Day222SiteICE3
RNA libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep with Sample Purification Beads (E7765, New England Biolabs, USA) following the manufacturer's instructions with the following options: 7 minutes fragmentation time, 5-fold Adapter Dilution, and seven cycles of final amplification. The libraries were run in a TapeStation 4150 with the D1000 Screentape and reagents to check for insert size distribution and the presence of adapter-primer dimers. After verification, the libraries were pooled equimolarly and sequenced on a NextSeq 500 (Illumina) using the 300 cycles, v2.5 (151bp paired-end sequencing) chemistry.