title: Nanopore sequencing of DRT2 RNP + 250 uM dNTP reaction
design: DNA was treated with RNase A, and ligated to a 5 pre-adenylated ssDNA adaptor ( /5rApp/CTGTCTCTTATACACATCTCCGAGCCCACGAGAC/3SpC3/) using thermostable  5 App DNA/RNA ligase (NEB). The reaction was treated with Proteinase K and products were purified with RNAclean XP beads. 0.5 uM of a ssDNA oligo complementary to the adaptor was added along with Phusion Flash master mix, and 8 cycles of polymerization were performed to allow second strand synthesis for any ssDNA in the reaction. Products were then purified and treated with the NEBNext Ultra II End Prep Repair/dA-tailing kit (NEB), purified with AMPure XP beads (Beckman Coulter) and ligated to the Oxford Nanopore Adaptor using the Ligation Sequencing Kit V14 (Oxford Nanopore). The library was loaded on a MinION Flow Cell (Oxford Nanopore) and sequenced for a total of 1,606,094 reads, with read polishing using Guppy.