Heavy Chain Isotype-Agnostic Immunoglobulin C gene RNA sequencing of mustela putorius furo: PBMCs
cDNA isolation: RNA was isolated from bulk cell samples using the RNeasy Mini Kit (Qiagen, 74104), and cDNA was synthesized using SuperScript IV (Invitrogen, 18091050), both per the manufacturers instructions. Priming strategy: a single forward primer targetting IGHJ genes (a sequence shared among all reported so far) combined with 3 prime RACE. Sequencing: performed in-house using Ligation Sequencing Kit V14 (Oxford NanoPore, SQK-LSK114), the NEB Next Companion Module (New England Biosciences, E718OS), Oxford Nanopore Technologys (ONT) MinION (FLO-MIN106D), and the Guppy Super Accurate basecaller (v4.3.4), followed by alignment using the Quickmerge software.