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Developing real-time PCR to identify cyanobacteria populations in lakes

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Isaac H. Stewart (Creator)
Institution
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/
Advisor
Parke Rublee

Abstract: The objective of this project was to develop and test primers for real-time PCR analysis of cyanobacteria. Primers were developed using an existing set of sequence libraries from the variable regions of the 16S rDNA for specific operational taxonomic units (OTUs) and then validated against existing clones and previously identified pure cultures. Environmental samples from High Point City Lake were then analyzed as a practical test to assess cyanobacterial abundance and growth patterns. Of 96 OTUs identified from the 883-sequence library, 30 OTUs were probed for in 18 genomic DNA samples collected from December 2007 through December 2008. OTU presence was confirmed and DNA was quantified from standard concentrations. OTUs were most abundant during summer months when water temperatures were highest. This study suggests that real-time PCR is an accurate way to monitor cyanobacteria diversity in City Lake. Given the conservation of 16S rDNA, climate change, increasing eutrophication of waterways, and general cyanobacteria abundance, this method of assessing cyanobacteria diversity is a step toward developing a rapid and efficient way to monitor the increasing abundance of cyanobacteria species and strains in water bodies.

Additional Information

Publication
Thesis
Language: English
Date: 2011
Keywords
Cyanobacteria, High Point City Lake, Real-Time PCR
Subjects
Cyanobacteria $x Ecology $z North Carolina
Cyanobacteria $x Identification
Polymerase chain reaction