Date of Award

Spring 5-7-2026

Document Type

Thesis

Degree Name

Bachelor of Science (BS)

Department

Marine Science

College

College of Science

First Advisor

Lauren M Stefaniak

Abstract/Description

Polymerase Chain Reaction (PCR) is a widely used molecular technique for amplifying specific regions of DNA, and its success depends heavily on the design of the primer pair being used. Primers must be specific enough to amplify the target region while minimizing the amplification of the non-target DNA, even more so when samples could contain DNA from multiple species. Cytochrome oxidase I (COI) is the standard mitochondrial gene used for DNA barcoding in animals, including ascidians. Several COI primer pairs have been used for ascidians, ranging from universally applicable primers to those intended to be ascidian specific. However, their breadth and specificity have yet to be systematically tested, bringing about the purpose of this study. This experiment will evaluate six COI primer pairs for breadth by comparing their amplification success across a diverse set of ascidian taxa and selectivity by comparing amplification success for non-ascidian invertebrates as well. Results from this project will help researchers select appropriate primers for ascidian barcoding studies and better understand the risk of amplifying non-target DNA in mixed samples.

Available for download on Monday, May 07, 2029

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