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Gentier Real-Time PCR System
3.3.4.4 High Resolution Melting
The melting temperature T
m
of dsDNA depends on its base sequence, fragment length and GC
content. In theory, any base change can cause difference in melting temperature; however, the
difference caused by a base change is tiny, usually less than 1 degree. As the resolution of
melting curve is not enough, it is required to read the fluorescence signal several times within
one degree of temperature change to accurately detect the difference of melting temperature.
Therefore, high resolution melting curve is often used for single nucleotide polymorphism (SNP)
analysis, mutation scanning, methylation research and genotyping, etc.
The high resolution melting analysis interface consists of seven functional modules:
Melting
Curve, Normalized Melting Curve, Diference Plot, Result Table and Sample Setting,
Normalized Peak Melting
and
Heat Map
. The high resolution melting analysis interface is
divided into four areas, which displays four functional modules (
Melting Curve, Normalized
Melting Curve, Diference Plot
and
Result Table
) by default, as shown in figure D 14.
Figure D 14. High Resolution Melting analysis interface
