Summarized experimental workflow. A Human liver total RNA sample was serially diluted from 1 to 1000 ng and coupled with a negative control of 0 ng of material. Libraries were prepared from each sample using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina with Unique Dual Index UMI Adapters RNA Set1. cDNA was selectively enriched with PCR containing unique dual indices (UDI) for multiplexing using 3 different numbers of PCR cycles adjusted according to the input amount. The native Illumina libraries were sequenced on two Illumina machines: NovaSeq 6000 and NovaSeq X (images obtained from https://www.illumina.com/ , Accessed 22.02.2025). The libraries underwent a conversion and then were also sequenced on AVITI (image obtained from https://www.elementbiosciences.com/products/aviti , Accessed 22.02.2025) and G4 (image obtained from https://singulargenomics.com/g4/ , accessed 22.02.2025).
