Genotyping of P450 CRISPR mutants in Heliconius erato using nanopore amplicaon sequencing

Heliconius erato eggs were injected with guides for that targeted three P450 genes (CYP40A5A4, HedCYP40A5A6a and HedCYP405A6b, the latter two were so similar in sequence that the same guide targeted both genes. Since all three CYP405A genes were close in sequence similarity with small exons and variable introns, primers were designed that amplified large regions of the genes (between 600bp and 1400bp) and then nanopore sequencing was used determine the sequence and check for deletions.