Ordered Two-Template Relay (OTTR) is a library preparation technique used to improve quantitation of highly modified non-coding RNA (ncRNA) species, which have been difficult to characterize using traditional cDNA sequencing approaches. OTTR leverages a retroelement reverse transcriptase (RT), termed BoMoC, with template jumping properties and high processivity across modified RNA templates, to generate cDNA products for next-generation sequencing (NGS). Overall, OTTR offers a streamlined approach for  cDNA library production of full-length and modified ncRNA targets.