(372k) Monitoring Algae Species in Bio-Fuel Production by CE-SSCP

Jernigan, A. C. - Presenter, University of Arkansas
Woods, L. - Presenter, University of Arkansas
Beitle, R. - Presenter, University of Arkansas
Hestekin, J. - Presenter, University of Arkansas
Hestekin, C. - Presenter, University of Arkansas

With its ability to use carbon dioxide from the atmosphere as a carbon source, and to clean nitrates and phosphorous from waste water, algae has the potential to be a significantly eco-friendly bio-fuel source. One of the problems faced in using algae as a bio-fuel is identifying and characterizing the best species for the purpose, and keeping them as the dominate species. The identification of algal communities can be accomplished by isolating DNA from water samples. These DNA samples can then be characterized by a technique called single-strand conformational polymorphism (SSCP) analysis of the 18S gene. The 18S gene is highly conserved among eukaryotes, but has unique small variable regions that can be used to differentiate between species. These regions can be amplified by polymerase chain reaction (PCR) giving small fragments of the gene sequence. SSCP, a method that is normally used to detect mutations, is the heat denaturing of these fragments resulting in single strand DNA. The refolding of the single strand DNA will form different secondary structures (conformations) depending on the nucleotide sequence. These different conformations can be separated due to their different electrophoretic mobility, even if they are the same size. The increased speed and automation of capillary electrophoresis (CE)-SSCP will allow for more rapid and reliable results allowing a quick and inexpensive method for monitoring the different algae species.