This protocol outlines the preparation of single-cell barcoded cDNA for Oxford Nanopore long-read sequencing. The input cDNA was initially barcoded using the Parse Biosciences Evercode platform for single-nucleus or single-cell RNA-seq (refer to the protocol “Evercode WT v2.2.1” at dx.doi.org/10.17504/protocols.io.eq2lyj9relx9/v1 or “Evercode WT Mega v2.2.1 at dx.doi.org/10.17504/protocols.io.8epv5xxrng1b/v1). Since we typically perform single-nucleus RNA-seq rather than single-cell RNA-seq, we enrich the cDNA for exome-containing fragments to reduce the number of intronic reads. This is achieved using Twist Biosciences exome panels and a modified version of the Parse Biosciences Gene Capture protocol (refer to the protocol “cDNA Exome Capture v1.0.1” at dx.doi.org/10.17504/protocols.io.36wgq3b83lk5/v1). The resulting product of this protocol is full-length, barcoded cDNA, excluding intron-only fragments, with Nanopore adapters added to both ends, creating a final library ready for sequencing. The first part of the protocol after setup, End Prep, involves using NEBNext reagents to repair the cDNA ends, resulting in 5′ phosphorylated, 3′ dA-tailed ends, followed by a bead-based cleanup. The next part, Adapter Ligation, adds Nanopore adapters to the end-prepped cDNA, followed by a bead-based cleanup and final elution. Please see the attachment for the original protocol.