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High-resolution DNA melting analysis of small amplicons is a simple and inexpensive technique for genotyping. Recent work within our lab studied the varying effects of amplicon size, melting rate and analysis methods. Two publications concluded that faster rates improve melting analysis, just like faster rates improve PCR.

Microfluidics allows precise and rapid control of temperature during melting. The resulting technique, High-Speed Melting Analysis (Pryor et al. Published in Clinical Chemistry), demonstrates high-speed melting reduces the time for melting analysis, decreases errors, and improves genotype discrimination of small amplicons. Combined with Extreme PCR, high-speed melting promises nucleic acid amplification and genotyping in < 1 min.

Microfluidic Instrument

High-Speed Melting can also be accomplished without microfluidics as described in Li. et al. (Published in Analytical Biochemistry), which describes the improved separation of homozygote and heterozygote (shown below), leading to easier identification (both qualitatively and quantitatively).

Figure: Genotype discrimination at slow (A and C) and fast (B and D) melting rates, either with (C and D) or without (A and B) curve overlay during analysis.(Li et al.)

Microfluidic Instrument

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