PAGE 7 OF 7 Nucleic Acids Research, 2010, Vol. 38, No. 11 e119 14. Osborne,A., Reis,A.H., Bach,L. and Wangh,L.J. (2009) Single-molecule LATE-PCR analysis of human mitochondrial genomic sequence variations. PLoS ONE, 4, e5636. 15. Santalucia,J. (1998) A uniﬁed view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics. Proc. Natl Acad. Sci. USA, 95, 1460–1465. 16. Le Novere,N. (2001) Melting, computing the melting temperature of nucleic acid duplex. Bioinformatics, 17, 1226–1227. 17. SantaLucia,J. (2007) Physical principles and visual-OMP software for optimal PCR design. Methods Mol. Biol., 402, 3–33. 18. Gillespie,D. (1997) Competitor primer asymmetric polymerase chain reaction. Patent No. 5627054. 19. Nordstro¨ m,T., Alderborn,A. and Nyre´ n,P. (2002) Method for one-step preparation of double-stranded DNA template applicable for use with PyrosequencingTM technology. J. Biochem. Biophys. Methods, 52, 71–82. 20. McKinzie,P.B. and Parsons,B.L. (2002) Detection of rare K-ras condon 12 mutations using allele-speciﬁc competitive blocker PCR. Mutat. Res., 517, 209–220. 21. Morlan,J., Baker,J. and Sinicropi,D. (2009) Mutation detection by real-time PCR: a simple, robust and highly selective method. PLoS ONE, 4, e4584.