We demonstrate that simple, non-invasive environmental DNA (eDNA) methods can detect transgenes of genetically modified (GM) animals from terrestrial and aquatic sources in invertebrate and vertebrate systems. We detected transgenic fragments between 82-234 bp through targeted PCR amplification of environmental DNA extracted from food media of GM fruit flies (Drosophila melanogaster), feces, urine, and saliva of GM laboratory mice (Mus musculus), and aquarium water of GM tetra fish (Gymnocorymbus ternetzi). With rapidly growing accessibility of genome-editing technologies such as CRISPR, the prevalence and diversity of GM animals will increase dramatically. GM animals have already been released into the wild with more releases planned in the future. eDNA methods have the potential to address the critical need for sensitive, accurate, and cost-effective detection and monitoring of GM animals and their transgenes in nature.

Bi-directional Sanger sequencing was conducted on an ABI 3730xl 96-capillary sequencer by the Centre d’expertise et de services Génome Québec. DNA sequences were aligned using BioEdit v.7.2.5 and ClustalW.

"F-", "GF-", "M-" files are raw transgene sequences from fruit fly, GloFish, and mouse respectively. "_final.fas" files are alignments of trimmed transgene sequences (three different fluorescent genes for GloFish).

ABI files can be opened through software such as FinchTV or any other DNA sequence electropherogram software.