Efficient RFLP-based Protocol for Routine Authentication of Drosophila

Authentication of strains is important for preventing genetic contamination before any experiment, which can compromise reproducibility and lead to misleading results. Here, we developed an approach that combines computational single nucleotide polymorphism (SNP) identification with molecular validation using restriction fragment length polymorphisms (RFLPs). This workflow enables rapid and precise confirmation of strains in an inexpensive, reproducible, and easily adaptable way for long-term stock maintenance across laboratories. We apply this protocol to <i>Drosophila melanogaster</i> from the Drosophila Genome Resource Panel (DGRP), which are commonly used in fruit fly research, providing a reliable context for ensuring the integrity of Drosophila genetic resources.