epdl amplicion sequencing of Paramormyrops sp. TEU
We designed primers to target all epdl2 paralogs in a given mormyrid species. As primer-binding regions, we chose the 5and 3 UTRs, close to the start and stop codons. We extracted DNA from the ethanol-preserved fin clips, from one individual of each of our studied species with the Qiagen DNeasy Blood & Tissue Kit.  Then we targeted all epdl2 paralogs for PCR amplification in 25 l reactions, with primer- and species-dependent reagents and conditions. The resulting amplicons were multiplexed and sequenced on an Oxford Nanopore Technologies (ONT) MinION device with a R9.4.1 flow cell (Nanopore sequencing). The multiplexed sequencing library was prepared with ONT kits: Native Barcoding Expansion 1-12 (cat. EXP-NBD104) & 13-24 (cat. EXP-NBD114), and Ligation Sequencing Kit (cat. SQK-LSK109), following manufacturers directions.  Reads were base-called and quality-filtered (Q > 7) with the high accuracy model from Guppy 4.2.3+f90bd04 (ONT), and barcoded reads passing filtering were demultiplexed with Guppy 4.4.1+1c81d62.