Marine Biology Commons^™

Reducing Host Dna Contamination In 16s Rrna Gene Surveys Of Anthozoan Microbiomes Using Pna Clamps, Alicia M. Reigel, Sarah M. Owens, Michael E. Hellberg

Faculty Publications

Efforts to study the microbial communities associated with corals can be limited by inefficiencies in the sequencing process due to high levels of host amplification by universal bacterial 16S rRNA gene primers. Here, we develop an inexpensive peptide nucleic acid (PNA) clamp that binds to a target sequence of host DNA during PCR and blocks amplification. We then test the ability of this PNA clamp to mitigate host contamination and increase overall microbial sequence coverage on samples from three coral species: the gorgoniansEunicea flexuosaandGorgonia ventalina,and the scleractinianPorites panamensis. The 20-bp PNA clamp was designed using DNA fromE. flexuosa. Adding the …

The Roles Of Host Species, Geographic Scale And Environmental Stressors In Shaping The Composition Of Coral Microbiomes, Alicia Marie Riegel Parker

LSU Doctoral Dissertations

Reef-building corals are long-lived and may take many centuries to adapt, making them especially susceptible to climate change. However, corals host microbial symbionts that can change quickly, potentially speeding acclimation. My dissertation aimed to determine the degree of coevolution and flexibility between corals and their microbiomes among hosts, across space, and in response to stress.

Microbial communities are usually surveyed by sequencing the 16S rRNA gene, however the PCR primers used also amplify coral DNA, thereby limiting prokaryotic read coverage. To mitigate this contamination, I designed a peptide nucleic acid clamp that increased the recovery of bacterial reads by 2-11x …

Fish Community Composition And Structure Near A Freshwater River Diversion In Southeastern Louisiana, Rachel L. Snider

LSU Master's Theses

Gulf of Mexico estuaries, particularly in Louisiana, are among the world’s most productive, with landings of commercially- and recreationally-important species exceeding all other contiguous US states. Coastal wetlands are disappearing at an alarming rate because they have been impounded from Mississippi River water and sediment. Although controversial, one proposed solution is to re-route the Mississippi River through diversions and siphons to supply the freshwater and sediments necessary to rebuild vanishing wetlands, particularly in Barataria Bay and Breton Sound, LA. This strategy is one approach outlined in the Louisiana Coastal Master Plan. This project aimed to describe the composition and structure …