Uli-epic: profiling RNA modifications from ultra-low input samples.

High-throughput sequencing with chemical labeling enables robust, transcriptome-wide detection of RNA modifications at single-nucleotide resolution. However, these methods typically require substantial RNA amounts due to harsh treatments. We introduce Uli-epic, an innovative library construction strategy that enables profiling epitranscriptomic modifications using 100 pg to 1 ng of RNA. Utilizing Uli-epic BID-seq, we investigate pseudouridine (Ψ) sites in neural stem cells and sperm RNA from wild-type and fetal growth restriction mice, using only 500 pg of rRNA-depleted RNA. Uli-epic GLORI quantifies m6A in sperm and neural stem cells from wild-type and fetal growth restriction mice, using 10 ng of rRNA-depleted RNA.