| Abstrakt: |
The Carolina Redhorse, Moxostoma sp. Carolina, is a member of the fish family Catostomidae and a threatened species in North Carolina, according to the North Carolina Wildlife Resources Commission. It is native to North Carolina and a small part of South Carolina and protected under the Endangered Species Act. Monitoring using a catch-and-release method, with PIT tagging (where small microchips are inserted under the scales to track movement) is not efficient and may be underrepresenting the species presence. This fish is localized to the Pee Dee River, and small sections of the Haw River, and Deep River in the Cape Fear River Basin. Environmental DNA (eDNA) may prove useful in monitoring the presence of species, both in waters where they have been previously recorded and have not yet been recorded. eDNA is any genetic material "shed" by an organism in its environment and has become more prevalent in aquatic species surveying. Species-specific PCR primers were designed using NCBI Blast, targeting a fraction of a Cytochrome b mitochondrial DNA gene. A map was created using previous surveys to assist in locating areas for initial sampling. Cartographical data is used to identify regions of interest where water samples are collected, filtered on-site, and processed through PCR and gel electrophoresis, using the primers designed. A positive result will show bands at lengths of 213 and 431 base pairs in the gel, and a negative result will show no banding, or banding at only one of the two target lengths. DNA isolated from a Carolina Redhorse fish sample will be used as a positive control and universal primers for GAPDH are used to confirm the presence of eDNA. Finally, sequencing will be used to validate the primers selected. [ABSTRACT FROM AUTHOR] |
