High-resolution melting

Manipulation of primer affinity improves high-resolution melting accuracy for imprinted genes

F. V. M. Rubatino, Carobin, N. V., Freitas, M. L., de Oliveira, V. T., Pietra, R. X., Oliveira, P. P. R., Bosco, A. A., and Jehee, F. S., Manipulation of primer affinity improves high-resolution melting accuracy for imprinted genes, vol. 14, pp. 7864-7872, 2015.

High-resolution melting (HRM) is considered an inexpensive, rapid, and attractive methodology for methylation analysis. In the application of the polymerase chain reaction (PCR) to methylation analysis, amplification efficiencies are biased towards unmethylated, rather than methylated, templates: a phenomenon known as PCR bias. To overcome PCR bias, primers that include CpG site(s) and are fully complementary to the methylated sequence have been proposed.

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