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 …