Genomic in vitro transcription and Nanopore direct RNA sequencing of a human B-Lymphocyte cell line.

Genomic DNA–templated in vitro transcribed RNA from a human B-lymphocyte cell line enables benchmarking Nanopore direct RNA sequencing modification detection using Dorado models.

• Why it matters: A true negative control is needed to benchmark RNA modification detection and quantify false positives for future Nanopore RNA modification model development.
• What they did: Dorado modification calling models were applied to Nanopore direct RNA sequencing data from IVT RNA made with genomic DNA from a human cell line, and 9-mer specific false-positive rates were computed for eight RNA modifications.
• The result: The study provides 9-mer specific false-positive rates for eight RNA modifications, establishing a quantitative comparison point for developing improved RNA modification models.