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Nucleic Acids Research, 2003, Vol. 31, No. 18 e114
© 2003 Oxford University Press

Ligation mediated PCR performed at low denaturation temperatures—PCR melting profiles

Aleksander Masny* and Andrzej Plucienniczak

Institute of Biotechnology and Antibiotics, Staroscinska 5, 02-516 Warsaw, Poland

*To whom correspondence should be addressed. Tel: +48 22 849 60 51; Fax: +48 22 849 33 32; Email: masnya{at}iba.waw.pl

We show that using low denaturation temperatures (80–88°C) during ligation mediated PCR (LM PCR) of bacterial DNA leads to the amplification of limited sets of the less stable DNA fragments. A set of electrophoretic patterns of such fragments obtained at different denaturation temperatures forms the PCR melting profile (PCR MP). A single pattern obtained for a given temperature and a set of patterns arising after application of several denaturation temperatures (PCR MP) are very specific for the given bacterial genome and may be used for strain characterisation and differentiation. The method may also be used for amplification and isolation of the less stable DNA fragments in a genome.


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