Date of Award

January 2017

Degree Type

Open Access Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

First Advisor

David M. Hayes

Second Advisor

Sherry L. Harrel

Third Advisor

Oliver R. Oakley

Abstract

Freshwater mussels have become some of the most imperiled species in North America and widespread populations have succumbed to pollution and many other anthropogenic-related factors. With molecular techniques evolving, a recent interest in ancient DNA and museum specimens has emerged and prompted a study to test the ability of several extraction methods to isolate DNA from museum mussel specimens. The purpose of this study was to determine if four DNA extraction methods had influence on total DNA yield (ng/mg) from mussel tissue. The hinge ligaments of freshwater mussels ranging in collection date (1984-2015) were used as the source of genetic material for this study. Additionally, collection date was tested for influence on the total DNA yield. An interaction between collection year and extraction method was also explored. A total of 40 hinge ligaments were removed from dried museum shells and subjected to four different DNA extraction methods. Total DNA yield (ng/mg) from the extractions was quantified using a Qubit 3.0 Fluorometer and a Nanodrop 2000. A modified CTAB extraction method was found to be statistically higher for extracting total DNA compared to the other three methods. This suggests that chloroform-based extractions may be optimal for DNA extraction from historic museum specimens containing fragile and degraded DNA. Future research will be necessary to determine the origin of DNA from the extracted genetic material. Now, with a more optimized extraction method, the hinge ligaments from dried shells stored museums can be used for extraction of host DNA and potentially eDNA released from other organisms.

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