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http://dx.doi.org/10.5657/FAS.2014.0339

Development of Clamping Probe for Rare DNA Detection using Universal Primers  

Kim, Meyong Il (Incheon Fisheries Research Institute)
Lee, Ki-Young (Department of Marine Biotechnology, Kunsan National University)
Cho, Sang-Man (Department of Aquaculture and Aquatic resources, Kunsan National University)
Publication Information
Fisheries and Aquatic Sciences / v.17, no.3, 2014 , pp. 339-344 More about this Journal
Abstract
PCR amplification with universal primer is a useful tool for speciation of symbionts in marine eukaryote coupled with robust separation method such as denaturing high performance chromatography (DHPLC). To overcome the biased amplification, clamping PCR is recommended to suppress the amplification of host gene. In this study, we evaluated the efficiency of rare gene detection for two kinds of clamping probes which were successfully utilized for eukaryotic symbiont analysis: C3 linked nucleotide (C3) and peptide nucleic acid (PNA). PNA was 3-4 orders of magnitude higher than that of C3 tested in clamping efficiency and rare gene detection. This represented that PNA could be a more competent clamping probe for the enhancement of PCR amplification for rare symbiont genes.
Keywords
Peptide nucleic acid (PNA); Clamping PCR; Symbiont; Clamping efficiency;
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