Application of Fluorescence Melting Curve Analysis for Dual DNA Detection Using Single Peptide Nucleic Acid Probe

Title
Application of Fluorescence Melting Curve Analysis for Dual DNA Detection Using Single Peptide Nucleic Acid Probe
Authors
황승용
Keywords
peptide nucleic acid; fluorescence melting curve analysis; DNA analysis genotyping
Issue Date
2015-05
Publisher
WILEY-BLACKWELL
Citation
BIOTECHNOLOGY PROGRESS, v. 31, NO 3, Page. 730-735
Abstract
Peptide nucleic acid (PNA) is an artificially synthesized polymer. PNA oligomers show greater specificity in binding to complementary DNAs. Using this PNA, fluorescence melting curve analysis (FMCA) for dual detection was established. Genomic DNA of Mycoplasma fermentans and Mycoplasma hyorhinis was used as a template DNA model. By using one PNA probe, M. fermentans and M. hyorhinis could be detected and distinguished simultaneously in a single tube. The developed PNA probe is a dual-labeled probe with fluorescence and quencher dye. The PNA probe perfectly matches the M. fermentans 16s rRNA gene, with a melting temperature of 72 degrees C. On the other hand, the developed PNA probe resulted in a mismatch with the 16s rRNA gene of M. hyorhinis, with a melting temperature of 44-45 degrees C. The melting temperature of M. hyorhinis was 27-28 degrees C lower than that of M. fermentans. Due to PNA's high specificity, this larger melting temperature gap is easy to create. FMCA using PNA offers an alternative method for specific DNA detection. (C) 2015 American Institute of Chemical Engineers
URI
http://onlinelibrary.wiley.com/doi/10.1002/btpr.2054/abstracthttp://hdl.handle.net/20.500.11754/24978
ISSN
8756-7938; 1520-6033
DOI
http://dx.doi.org/10.1002/btpr.2054
Appears in Collections:
GRADUATE SCHOOL[S](대학원) > BIONANOTECHNOLOGY(바이오나노학과) > Articles
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