Evaluating Manta Ray Mucus as an Alternative Dna Source for Population Genetics Study: Underwater-sampling, Dry-storage and Pcr Success Advanced Genetic Analyses Can Contribute Relevant Information on Effective Population Size

View the peer-reviewed version (peerj.com/articles/1188), which is the preferred citable publication unless you specifically need to cite this preprint. manta ray mucus as an alternative DNA source for population genetics study: underwater-sampling, dry-storage and PCR success. PeerJ 3:e1188 Abstract 13 Sharks and rays are increasingly being identified as high-risk species for extinction, 14 prompting urgent assessments of their local or regional populations. 16 and connectivity among populations although acquiring sufficient regional sample sizes can 17 be challenging. DNA is typically amplified from tissue samples which are collected by hand 18 spears with modified biopsy punch tips. This technique is not always popular due mainly to a 19 perception that invasive sampling might harm the rays, change their behaviour, or have a 20 negative impact on tourism. To explore alternative methods, we evaluated the yields and PCR 21 success of DNA template prepared from manta ray mucus collected underwater and captured 22 and stored on a Whatman FTA TM Elute card. The pilot study demonstrated that mucus can be 23 effectively collected underwater using a toothbrush. DNA stored on cards was found to be 24 reliable for PCR-based population genetic studies. We successfully amplified mtDNA ND5, 25 nuclear DNA RAG1, and microsatellite loci for all samples. As the yields of DNA with the 26 tested method were low, further improvements are desirable for assays that may require larger 27 amounts of DNA, such as population genomic studies using emerging next-gen sequencing.