부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ab.2005.04.009 Byline: C. Peano (a), F. Lesignoli (b), M. Gulli (a), R. Corradini (b), M.C. Samson (a), R. Marchelli (b), N. Marmiroli (a) Keywords: PCR clamping; PNA; GMO traceability; Maize; Soybean Abstract: In the present study a peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method was developed and applied to the detection of genetically modified organisms (GMO), to test PCR products for band identity and to obtain a semiquantitative evaluation of GMO content. The minimal concentration of PNA necessary to block the PCR was determined by comparing PCRs containing a constant amount of DNA in the presence of increasing concentration of target-specific PNA. The lowest PNA concentration at which specific inhibition took place, by the inhibition of primer extension and/or steric hindrance, was the most efficient condition. Optimization of PCR clamping by PNA was observed by testing five different PNAs with a minimum of 13bp to a maximum of 15bp, designed on the target sequence of Roundup Ready soybean. The results obtained on the DNA extracted from Roundup Ready soybean standard flour were verified also on DNA extracted from standard flours of maize GA21, Bt176, Bt11, and MON810. A correlation between the PNA concentration necessary for inducing PCR clamping and the percentage of the GMO target sequence in the sample was found. Author Affiliation: (a) Department of Environmental Sciences, Division of Genetics and Environmental Biotechnology, University of Parma, 43100, Parma, Italy (b) Department of Organic and Industrial Chemistry, University of Parma, 43100, Parma, Italy Article History: Received 29 December 2004