Direct RNA sequencing of Human rRNA
Nanopore direct RNA sequencing for rRNA was performed as previously described6. Total RNA was isolated from BE(2)-C control, BE(2)-C shDKC1 738, BE(2)-C shPUS7-33, 293-FT ASO control, and 293-FT ASO SNORA41 cell lines. Total RNA was enriched for rRNA by depleting mRNA using poly(A) enrichment. Following rRNA enrichment, a poly-A tail was added using a poly(A) Tailing Kit (NEB, E7490S). The direct RNA sequencing library was prepared according to the standard protocol and library loaded into a Flongle flow cell for sequencing using MinKnow (version 23). Each flow cell was sequenced for 24 hours.